KR20070023670A - Oxidation by hydrogen peroxide - Google Patents

Abstract

The present invention is catalyzed by a monooxygenase enzyme and generates hydrogen peroxide at a rate below the rate at which hydrogen peroxide is used in the reaction simultaneously with an oxidation reaction catalyzed by a monooxygenase enzyme and using hydrogen peroxide as the oxidant. oxidative damage in use, and the mono oxide dioxygenase claim as an oxidizing agent is the presence of a method of performing the oxidation reaction at a low level or H ₂ O ₂ or hydroxyl radical isolated H ₂ O ₂ or hydroxyl radical, which control the concentration of the It provides a method of carrying out the reaction.

Monooxygenase enzyme, oxidation reaction, hydrogen peroxide, hydroxyl radical, oxidative damage

Description

Oxidation by Hydrogen Peroxide {OXIDATION BY HYDROGEN PEROXIDE}

The present invention relates to a method of carrying out an oxidation reaction.

Monooxygenase enzymes catalyze the oxidation of a very broad range of substrates. In general, co-factors and one or more electron-transfer partner proteins (reductases) are required for the monooxygenase enzyme to catalyze this reaction. However, monooxygenase enzymes can utilize hydrogen peroxide (H ₂ O ₂ ) as the oxidant because H ₂ O ₂ acts as a source of dioxygen and two electrons. The use of H ₂ O ₂ to advance the oxidation reaction is known as “peroxide shunt”.

Summary of the Invention

Monooxygenase enzymes generally have a higher K _m for H ₂ O ₂ compared to peroxidase enzymes (eg about 20 mM). Therefore, high levels of H ₂ O ₂ are required for significant levels of monooxygenase enzyme activity when performing oxidation reactions with peroxide shunts. For example, the initial rate of monooxygenase activity using 50 mM H ₂ O ₂ is much lower than when the natural cofactor NAD (P) H is used with a physiological electron-transfer partner.

The present invention reduces the oxidative damage caused to the monooxygenase enzyme by allowing the oxidation reaction with the peroxide shunt to be carried out so that an excessive level of H ₂ O ₂ is not present, thereby more efficiently oxidizing the reaction with the peroxide shunt. Provides a way to do it.

As the mono-oxide is H ₂ O ₂ used in the oxidation reaction catalyzed by a dioxygenase claim this is that at the same time the H ₂ O ₂ generated below this H ₂ using a rate of O ₂ to improve the efficiency of oxidation reaction and increase product yields . Various methods, such as electrochemical reactions, the use of enzymes or precursors, can be used to produce H ₂ O ₂ at the required rate.

Accordingly, the present invention is catalyzed by a monooxygenase enzyme, which comprises producing a hydrogen peroxide at or below the rate at which hydrogen peroxide is used in the reaction simultaneously with an oxidation reaction catalyzed by a monooxygenase enzyme and using hydrogen peroxide as the oxidant, Hydrogen peroxide is used as the oxidant and the oxidative damage of monooxygenases provides a way to carry out oxidation reactions that occur at low levels.

The present invention also relates to mono-oxide dioxygenase is catalyzed by the enzyme performs an oxidation reaction using hydrogen peroxide as an oxidizing agent in the presence of H ₂ O ₂ or hydroxyl radical isolated H ₂ O ₂ or hydroxyl radical, which controls the concentration of the Which comprises a catalyst catalyzed by a monooxygenase enzyme and utilizing hydrogen peroxide as the oxidant and oxidative damage of the monooxygenase occurs at low levels.

Sequence description

SEQ ID NO: 1 shows the nucleotide sequence of cytochrome P450 _Cam from Pseudomonas putida .

SEQ ID NO: 2 shows the amino acid sequence of cytochrome P450 _Cam from Pseudomonas putida.

SEQ ID NO: 3 shows the nucleotide sequence of cytochrome P450 _{BM- 3} from Bacillus megaterium .

SEQ ID NO: 4 shows the amino acid sequence of cytochrome P450 _{BM- 3} from Bacillus megaterium. The first 472 amino acid residues form the heme domain. The last 585 amino acid residues form a reductase domain. All 1048 amino acid residues form a homoenzyme.

For convenience herein we employ one of skill in the art to indicate mutants by listing the amino acid present at that position in the mutant following the original amino acid residue present at any position in the sequence, for example F87 is located in the wild-type sequence. Refers to phenylalanine of 87, F87A indicates that the phenylalanine at position 87 in the wild-type sequence changed to alanine in the variant. Numbers assigned to amino acid residues begin with the amino acid following the starting methionine residue.

The mutants used in the examples were F87A (single mutation; SEQ ID NO: 5 and SEQ ID NO: 6) and F87V L188Q A74G (triple mutation; SEQ ID NO: 7 and SEQ ID NO: 8).

SEQ ID NO: 5 shows the amino acid sequence of the F87A mutant of cytochrome P450 _{BM- 3} from Bacillus megaterium.

SEQ ID NO: 6 shows the nucleotide sequence of the F87A mutant of cytochrome P450 _{BM- 3} derived from Bacillus megaterium.

SEQ ID NO: 7 shows the amino acid sequence of the F87V L188Q A74G mutant of cytochrome P450 _{BM- 3} derived from Bacillus megaterium.

SEQ ID NO: 8 shows the nucleotide sequence of the F87V L188Q A74G mutant of cytochrome P450 _{BM- 3} derived from Bacillus megaterium.

SEQ ID NO: 9 shows the nucleotide sequence of B-276 alkene epoxidase subunit 1 derived from Nocardia coralline .

SEQ ID NO: 10 shows the amino acid sequence of B-276 alkene epoxidase subunit 1 derived from Nocardia Coralline.

SEQ ID NO: 11 shows the nucleotide sequence of B-276 alkene epoxidase subunit 2 derived from Nocardia coralline.

SEQ ID NO: 12 shows the amino acid sequence of B-276 alkene epoxidase subunit 2 derived from Nocardia Coralline.

SEQ ID NO: 13 bakta Xanthomonas species (Xanthobacta sp . ) Shows the nucleotide sequence of the Py2 alkene monooxygenase alpha subunit.

SEQ ID NO: 14 shows the amino acid sequence of the Py2 alkene monooxygenase alpha subunit from Xanthobacta spp.

SEQ ID NO: 15 shows the nucleotide sequence of a Py2 alkene monooxygenase beta subunit derived from Xanthobta spp.

SEQ ID NO: 16 shows the amino acid sequence of the Py2 alkene monooxygenase beta subunit derived from Xanthobta spp.

SEQ ID NO: 17 shows the nucleotide sequence of a Py2 alkene monooxygenase gamma subunit derived from Xanthobta spp.

SEQ ID NO: 18 shows the amino acid sequence of the Py2 alkene monooxygenase gamma subunit derived from Xanthobacta spp.

SEQ ID NO: 19 shows the nucleotide sequence of a soluble methane monooxygenase alpha subunit derived from Methylococcus capsulatas .

SEQ ID NO: 20 shows the amino acid sequence of the soluble methane monooxygenase alpha subunit derived from Methylcococcus capellatas.

SEQ ID NO: 21 shows the nucleotide sequence of the soluble methane monooxygenase beta subunit derived from Methylcococcus capellatas.

SEQ ID NO: 22 shows the amino acid sequence of the soluble methane monooxygenase beta subunit derived from Methylcoccus capellatas.

SEQ ID NO: 23 shows the nucleotide sequence of the soluble methane monooxygenase gamma subunit derived from Methylcococcus capellatas.

SEQ ID NO: 24 shows the amino acid sequence of the soluble methane monooxygenase gamma subunit derived from Methylcococcus capellatas.

SEQ ID NO: 25 shows the nucleotide sequence of GPol alkane hydroxylase ( Alk B gene) derived from Pseudomonas oleovorans .

SEQ ID NO: 26 shows the amino acid sequence of GPol alkane hydroxylase from Pseudomonas oleoboranth.

SEQ ID NO: 27 shows the nucleotide sequence of a toluene 2-monoxyoxygenase alpha subunit derived from Burkholderia cepacia .

SEQ ID NO: 28 shows the amino acid sequence of the toluene 2-monooxygenase alpha subunit from Burkholderia Sephacia.

SEQ ID NO: 29 shows the nucleotide sequence of the toluene 2-monooxygenase beta subunit derived from Burkholderia Sephacia.

SEQ ID NO: 30 shows the amino acid sequence of the toluene 2-monooxygenase beta subunit derived from Burkholderia Sephacia.

SEQ ID NO: 31 shows the nucleotide sequence of a toluene 2-monooxygenase gamma subunit from Burkholderia Sephacia.

SEQ ID NO: 32 shows amino acid sequence of toluene 2-monooxygenase gamma subunit from Burkholderia Sephacia.

SEQ ID NO: 33 shows the nucleotide sequence of a phenol hydroxylase (pheA) gene derived from Bacillus stearothermophilus .

SEQ ID NO: 34 shows the amino acid sequence of the phenolic hydroxylase gene derived from Bacillus stearothermophilus.

SEQ ID NO: 35 shows the nucleotide sequence of stearoyl-ACP desaturase derived from Helianthus annuus .

SEQ ID NO: 36 shows the amino acid sequence of stearoyl-ACP desaturase from Helianthus annuus.

It should be understood that the present invention is not limited to the specific embodiments. It should also be understood that various ways of applying the disclosed methods may be modified to suit specific needs in the art. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments of the invention only, and is not intended to be limiting of the invention.

Also, as used in the description and claims of the present specification, the singular forms “a,” “an,” and “the” include plural forms unless the context clearly dictates otherwise. Thus, for example, reference to "substrate" includes two or more substrates, and reference to "enzyme" includes two or more enzymes.

All publications, patent documents, and patent applications mentioned herein, either above or below, are hereby incorporated by reference in their entirety.

The method of the present invention can oxidize various substrates. Such substrates include, but are not limited to, alkanes, aromatics, terpenoids, alkenes and fatty acids.

Suitable alkanes include, but are not limited to, methane, ethane, propane, butane, pentane, hexane, heptane, n-octane, n-nonane, n-decane, n-dodecane and n-hexadecane. When alkanes are oxidized, alcohol is produced. Oxidation of methane to methanol is of great technical and economic importance. Long chain alcohols (eg n-dodecanol) are used for the synthesis of fatty acid derivatives and heavy chain alcohols (eg n-octanol) are synthetic intermediates.

Suitable aromatic compounds include, but are not limited to, benzene, toluene, xylene, chlorobenzene, phenols and their substituents and the like. Phenolic and catechol-based products are used for the synthesis of aromatic and flavored compounds.

Suitable terpenoid compounds include, but are not limited to, monoterpenes such as limonene, pinene, terpinene and ocymen, sesquiterpenes such as valentene and aromadendrene and triterpenes such as steroidal compounds It is not limited. The product is a synthetic intermediate of precise aromatic and flavoring chemicals and pharmaceuticals.

Suitable alkenes include, but are not limited to, simple molecules such as propene, hex-1-ene, hex-2-ene and styrene, and compounds having carbon-carbon double bonds in complex molecules. It is very important for synthesis that the alkene is selectively epoxidized to become a single enantiomer. Optically pure propene oxide and styrene oxide are very useful intermediates in the synthesis.

Hydroxylated fatty acids are precursors to polymers.

Monooxygenase enzyme

The enzyme used to carry out the oxidation reaction according to the invention is a monooxygenase enzyme. One skilled in the art can determine whether a given enzyme is a monooxygenase enzyme using standard techniques in the art. Typically, in particular progesterone clusters of heme-free iron enzymes can be characterized using protein crystallography, since they generally do not possess chromophores. Otherwise, those skilled in the art will typically use sequence alignment, iron content and subunit composition to find conserved motifs such as active sites.

The K _m of the monooxygenase enzyme for H ₂ O ₂ is preferably at least 15 nM, at least 20 nM, at least 25 nM, at least 30 nM, at least 35 nM, at least 40 nM, at least 45 nM, or at least 50 nM.

Examples of monooxygenase enzymes include, but are not limited to, cytochrome P450 monooxygenase and heme-free two-iron monooxygenase enzymes. Suitable heme-free two-iron monooxygenase enzymes include methane monooxygenase (Colby et al., Biochem. J., 1977; 165: 395-402), Dalton, Adv. Appl. Microbiol , 1980; 26: 71-87, Fox et al., J. Biol. Chem., 1989; 264: 10023-10033, Fox et al., Methods Enzymol., 1990; 188: 191-202. McDonald et al., Appl. Environ.Microbiol., 1997; 63: 1898-1904), alkanes hydroxylases [van Beilen et al., Enzyme Microb. Technol., 1994; 16: 904-911, toluene monooxygenase (Luykx et al., Biochem. Biophys. Res. Commun., 2003; 312: 373-379], Pikus et al., Biochemistry, 1996; 35: 9106 -9119, Newman & Wackett, Biochemistry, 1995; 34: 14066-14076), alkene monooxygenase (Gallagher et al., Eur. J. Biochem., 1997; 247: 635-641), [ Lange & Que, Curr. Opin. Chem. Biol., 1998; 2: 159-172], Zhou et al., FEBS Lett., 1998; 430: 181-185], phenol monooxygenase ([Divari] et al., Eur. J. Biochem., 2003; 270: 2244-2253] and steroid desaturases [Shanklin et al., Biochemistry, 1994; 33: 12787-12794], and the like. The free two-iron monooxygenase enzyme is typically of eukaryotic or prokaryotic origin, preferably of bacterial, fungal, yeast, plant or animal origin The preferred sequences are shown in SEQ ID NOs: 1-36.

The enzyme used in the method of the present invention is typically a cytochrome P450 enzyme of eukaryotic or prokaryotic origin. Typically, the cytochrome P450 monooxygenase is characterized by a 446-450 nm heme Soret band for the ferrous-carbon monoxide complex. The enzyme may be derived from bacteria of the genus Pseudomonas because of its bacterial, fungal, yeast, plant or animal origin. The enzyme is the naturally occurring form of P450, e.g., Bacillus MEGATHERIUM origin of the _cam P450, P450 _{BM -3,} a P450 _terp, saccharide of Pseudomonas species-derived u Fora Erie Tra Ke (Saccharopollyspora erythraea) derived P450 _eryF and also P450 105 D1 (CYP105) from Streptomyces griseus strain.

Alternatively, the enzyme may be a mutant of P450 in naturally occurring form. The mutant retains the intrinsic biological activity of naturally occurring enzymes, ie the catalytic ability of oxidation reactions using H ₂ O ₂ . The mutant may have one or more mutations in its enzymatic active site.

An amino acid "in the active site" is an amino acid that identifies or defines the site to which the substrate binds during catalysis, or an amino acid that identifies or defines the site that must pass before the substrate reaches the catalytic site. Thus, these amino acids typically interact with the substrate during entry into the catalytic site or during catalysis. This interaction typically occurs through electrostatic interaction forces (between charged or polar groups), hydrophobic interaction forces, hydrogen bonding forces or van der Waals forces.

Amino acids in the active site can be identified by methods conventional to those skilled in the art. Such methods include labeling studies in which an enzyme is coupled to a substrate, wherein the substrate modifies ('labels') the amino acid in contact with it. Alternatively, the crystal structure of the enzyme to which the substrate is bound may be found to estimate amino acids in the active site.

Monooxygenase enzymes may have one, two, three, four, five to ten, ten to twenty or more other mutations, such as substitutions, insertions, or deletions. Amino acid substitutions can occur in the amino acid sequence of a naturally occurring enzyme, for example one, two, three, four or five to ten, twenty or thirty substitutions can occur. For example, conservative substitutions may occur, such as in Table 1 below. The amino acids listed in the same column of the second column in Table 1, preferably those listed in the same column of the third column, may be substituted for one another:

Conservative Amino Acid Substitutions Non-aromatic Nonpolar G, A, P I, L, V Polarity-non-charge C, S, T, M N, Q Polarity-charge D, E H, K, R Aromatic H, F, W, Y

Mutations can be present within the active site or outside the active site. Typically, mutations are present in “second sphere” residues that affect or contact the position or orientation of one or more amino acids present in the active site. Since the insertion typically occurs at the N and / or C terminus, the enzyme may be part of a chimeric protein. Deletion typically includes deletions of amino acids not involved in catalysis, for example amino acids outside the active site (thus the enzyme is a mutated fragment of a naturally occurring enzyme). Thus, the monooxygenase enzyme may comprise only amino acids required for oxidative activity.

Typically, mutations in the active site alter the position and / or shape of the substrate when it is bound to the active site. Mutations can make it easier for the site to be oxidized to access the heme group. Thus, a mutation can be a substitution with a smaller or larger amino acid, or with an amino acid having a more polar or less polar side chain.

Mutations typically increase protein stability or make protein purification easier. Typically, these typically prevent dimerization of the protein by removing cysteine residues from the protein (e.g., position 334 of P450 _cam , or by replacing cysteine, preferably with alanine, at the equivalent position of the homologue). . These typically introduce a deletion or poly-histidine tag, or mutate the N-terminal membrane anchoring sequence, for example, so that the protein is made in soluble form. Mutations typically inhibit protein oligomerization, eg, oligomerization due to contact between hydrophobic patches present on the protein surface.

Mutations can affect the way the enzyme uses H ₂ O ₂ so that the efficiency of the reaction is improved. For example, replacing all methionine residues in the heme domain of P450 _{BM- 3} with norleucine results in a two-fold increase in the peroxygenase activity of the enzyme [Cirino et al., Biotechnol. Bioeng., 2003; 83 (6): 729-734]. In addition, direct evolutionary studies have been reported to find mutant enzymes that are more resistant to peroxides [Cirino & Arnold, Angew. Chem. Int. Ed., 2003; 42: 3299-3301.

Thus, mutant enzymes are typically at least 70% homologous to naturally occurring enzymes based on amino acid identity.

The mutant protein referred to herein (ie, the protein described as a mutant of another protein) is at least 20, preferably at least 30, for example 40, 60 or 100 or more of the protein involved. It is typically at least 70% homologous over, or at least 80% or 90%, more preferably at least 95%, at least 97% or at least 99% homologous to adjacent amino acids. Contiguous amino acids may comprise an active site. Alternatively, this homology can be determined not only for adjacent amino acids but also for amino acids in the active site.

Homology can be determined using known methods. For example, the UWGCG Package provides a BESTFIT program that can be used for homology calculations (eg used in its default settings) [Devereux et al (1984) Nucleic Acids Research 12, p387-395]. PILEUP and BLAST, as described, for example, in Altschul SF (1993) J Mol Evol 36: 290-300, Altschul, S, F et al (1990) J Mol Biol 215: 403-10. Algorithms can be used for homology calculations or sequence alignments (typically used for their default settings).

Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information (NCBI) (http://www.ncbi.nlm.nih.gov/). This algorithm verifies that a short word of length W of the sequence of interest meets or matches a slightly positive threshold score T when aligned with the same word length in the database sequence, thereby matching a high score sequence pair (HSP, High Scoring Sequence Pair). T is referred to the neighboring word score threshold (Altschul et al, supra). This starting neighboring word hit serves as the basis for the initial search to find the HSP containing it. The word hits extend in both directions along each sequence as long as the cumulative alignment score can be increased. The extension for the word hit in each direction is when the cumulative alignment score falls by an amount X from its maximum reached value; When the cumulative score drops below zero due to accumulation of one or more negative-score residue alignments; Or stops when the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLAST program defaults to the word length (W) 11, BLOSUM62 score matrix (see Henikoff and Henikoff (1992) Proc. Natl. Acad. Sci. USA 89: 10915-10919) alignment (B) 50, expected (E ) 10, M = 5, N = 4 and the comparison of both strands is used.

The BLAST algorithm performs a statistical analysis of the similarity between the two sequences (see, eg, Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90: 5873-5787). One of the similarity measures provided by the BLAST algorithm is the minimum total probability (P (N)), which provides an indication of the probability by which two nucleotide sequences or two amino acid sequences coincide. For example, when the minimum total probability when comparing the first sequence to the second sequence is less than about 1, preferably less than about 0.1, more preferably less than about 0.01, most preferably less than about 0.001, these sequences are mutually Considered similar.

Mutants include fragments of the above-mentioned sequences. Such fragments retain monooxygenase activity. The fragment may be at least 300, at least 400, or at least 450 amino acids in length. Such fragments can be used to generate chimeric enzymes, which will be described in more detail below.

Mutants also include chimeric proteins comprising fragments or portions of naturally occurring enzymes. One or more amino acids may alternatively or additionally be added to the polypeptides described above. The N-terminus or C-terminus of a naturally occurring enzyme or variant or fragment thereof may be extended. This extension may be very short, for example 1 to 10 amino acids in length. Alternatively, the extension may be longer. Carrier proteins may be fused to the amino acid sequences described above. Thus, fusion proteins incorporating one of the enzymes described above can be used in the present invention.

Naturally occurring enzymes or mutants thereof may be chemically modified. Numerous side chain modifications are known in the art and can be applied to the side chains of the enzymes discussed above. Examples of such modifications include glycosylation, phosphorylation, reductive alkylation by reaction with aldehydes and subsequent reduction with NaBH ₄ , amidation with methylacetimidadate or amino acid modification with acylation with acetic anhydride. . It is preferred that the modification is glycosylation.

In general, the mutations discussed herein are carried out using methods known in the art, such as site-directed mutagenesis of enzymes, PCR and gene shuffling methods, or in the cycle of site-directed mutagenesis. It is introduced into the enzyme by using various mutagenic oligonucleotides. Thus, mutations can be introduced in a designated or random manner. Thus, mutagenesis produces one or more polynucleotides encoding one or more different mutants. Typically. A library of mutant oligonucleotides is generated, which can be used to generate a library of mutant enzymes.

Enzymes can be prepared via synthetic or recombinant means using methods known in the art. The amino acid sequence of the monooxygenase enzyme may be modified to include non-naturally occurring amino acids or to increase enzyme stability. If the enzyme is prepared by synthetic means, these amino acids may be introduced during the preparation process. The protein or peptide may be modified after synthesis or recombinant production.

Enzymes may also be prepared using D-amino acids. In this case, the amino acids will be linked to the reverse sequence in C to N orientation. This is common in the art for preparing such proteins or peptides.

Enzyme is derived from the recombinant expression vector in situ ) can be produced in cells via expression. If desired, the expression vector carries an inducible promoter that controls the expression of the polypeptide. The enzyme can be produced on a large scale by purification with any protein liquid chromatography system after recombinant expression. Preferred protein liquid chromatography systems include FPLC, AKTA systems, Bio-Cad systems, Bio-Rad BioLogic systems, and Gilson HPLC systems.

Oxidation reaction

The process of the invention relates to carrying out a high efficiency oxidation reaction catalyzed by a monooxygenase enzyme. Highly efficient oxidation reactions are reactions that occur without significant turnaround in enzyme turnover or product yield or inactivation of monooxygenase enzymes. Preferably, the monooxygenase enzyme is at least 70%, at least 80%, at least 90% of the activity exhibited at the start of the reaction after 1 hour, 2 hours, 6 hours, 12 hours, 1 day, 2 days or 5 days. , 95% or more or 100%.

Typically, the methods of the present invention are performed in vitro, such as in cell-free systems and the like.

The reaction proceeds by "peroxide shunt". The reaction of the present invention is carried out in the presence of monooxygenase enzyme (a), substrate (b) and H ₂ O ₂ (c). The reaction is typically carried out under aerobic conditions and no cofactor is required. The production of (c) will be discussed in more detail below. In this system, the flow of electrons is typically (c) → (a) → (b).

In this method, the concentrations of (a) and (b) are typically 10 ^-8 to 10 ^-2 M, preferably 10 ^-6 to 10 ^-4 M. Typically, the concentration ratio of (a) :( b) is 0.1: 10 to 1:10, preferably 1: 0.5 to 1: 2 or 1: 0.8 to 1: 1.2. It is preferable that the density | concentration of (b) is higher than the density | concentration of (a). Preferred concentrations of (a) are such that sufficient products are produced to be detected by available assays such as GC, HPLC and the like upon reaction with the substrate. This is typically at the level of μM amounts.

Generally, the method is carried out at a temperature and / or pH at which the monooxygenase enzyme is functional, for example at a temperature and / or pH at which the enzyme has at least 20%, at least 50%, at least 80% of its highest activity. Typically, the pH is 2 to 11, for example 5 to 9 or 6 to 8, preferably 7 to 7.8 or 7.4. The pH may be maintained using a system based on a suitable buffer such as phosphate or acetate. Typically, the temperature is 0 to 80 ° C, for example 25 to 75 ° C, 30 to 60 ° C or 50 to 80 ° C. It is preferable that the said temperature is 20-40 degreeC.

Typically, at least 20 turnovers occur per minute in the process, for example at least 50, at least 100, at least 200, at least 300, at least 500, at least 500 (where the turnover is the nanomolarity of the product formed per nM of enzyme ( measured in nM)).

Typically, the production rate of H ₂ O ₂ is 1 μg, 2 μg or 3 μg or less per minute per mg of monooxygenase enzyme. Typically, the concentration of H ₂ O ₂ is no more than 0.1 mM, 0.5 mM or 1 mM throughout the reaction. Typically, the reaction lasts at least 60 minutes, at least 240 minutes, at least 6 hours or at least 10 hours.

The process of the present invention can be carried out in a substrate where the monooxygenase substrate is liquid under reaction conditions. The process of the invention may be carried out in a solvent. Suitable solvents include, but are not limited to, water / organic and aqueous buffer / organic solvent systems mixed with aqueous buffer solutions. The organic solvent is preferably a hydrocarbon such as hexane, benzene, acetonitrile, lower aliphatic alcohols, ketones and dioxanes, dimethylformamide and dimethyl sulfoxide and mixtures thereof. The solvent is typically one in which the reagents and products are highly soluble and the stability and activity of the monooxygenase enzyme are maintained.

The reaction can be carried out in a homogeneous system containing all components in solution. Typically, the monooxygenase enzyme and the substrate are mixed together in a suitable solvent in a stirred tank reactor and the reaction is carried out batchwise, semi-batch or continuously.

Alternatively, the monooxygenase enzyme can be immobilized on a suitable solid support, for example silica, before carrying out the method of the invention. The immobilized monooxygenase enzyme can be charged to a fixed bed reactor and the substrate will pass through this enzyme. In one embodiment, enzymes that produce H ₂ O ₂ (discussed in more detail below) can be immobilized as monooxygenase enzymes on the same or different materials. Procedures for immobilizing enzymes are known in the art. Examples of such procedures include, but are not limited to, covalent coupling with insoluble organic or inorganic supports, capture in gels and adsorption to ion exchange resins or other adsorbents [GF Bickerstaff ed., "Immobilization of Enzymes and Cells". , "Humana Press, Totowa. New Jersey, 1997].

In a further embodiment, the hydrophilic membrane on the "exhaust" side flows the hydrophilic compound to the "product" side of the flow reaction cell after the membrane on the "entrance" side slowly receives the substrate from the "reactant" side. In this case, H ₂ O ₂ can be produced outside the membrane and flow through the membrane to a mobile or fixed enzyme.

In one embodiment, H ₂ O ₂ is preferably produced by one of the methods discussed in more detail below. In another embodiment, H ₂ O ₂ or hydroxyl radical sequestrants are used to segregate excess H ₂ O ₂ or hydroxyl radicals during the oxidation reaction. The sequestering agent may be a chelating agent. In one embodiment, the chelating agent is EDTA. EDTA inhibits the production of hydroxyl radicals, for example produced by the reaction of trace iron (or copper) with H ₂ O ₂ .

By electrochemical reaction H ₂ O ₂ Generation of

In the process of the invention H ₂ O ₂ may be produced by an electrochemical reaction. In general, the electrochemical reaction is a means of introducing a current into a liquid, preferably a solution. Typically, an electrochemical reaction is an oxidation or reduction reaction that occurs at an electrode through which a current flows. The electrode is a solid capable of conducting electricity, typically a carbon-based or metallic solid, which is connected to an external source or sink that is in contact with the liquid, preferably the solution. The electrode may be positively charged (cathode) or negatively charged (anode). Two or more electrodes can form an electrochemical cell from which an external wire can be connected from each electrode to an external electrical device. The oxidation reaction or the reduction reaction takes place at one electrode, but the redox reaction may take place in the electrochemical cell or directly in the liquid.

The production of H ₂ O ₂ using electrochemical reactions is energy efficient. H ₂ O ₂ is typically produced while controlling oxygen molecules to be electrochemically reduced to hydrogen peroxide. The surface area and overvoltage of the cathode is a key consideration in the two-electron reduction of oxygen molecules to hydrogen peroxide. Typically, carbon-based cathodes are used and these can be transformed into compounds known to lower the overvoltage for this reaction. Electrode materials and variants for performing this operation effectively and efficiently are known in the art. Typically, the reduction of O ₂ and thus the production of hydrogen peroxide can be controlled by the voltage applied to the cathode. The voltage applied to the cathode will depend on any modifications to the cathode and the manufactured cathode.

The electrochemical reaction used in the process of the invention may be sonoelectrochemical reduction of dioxygen. This method is known in the art [Compton et al., Electroanalysis, 1997; 9 (7): 509-522.

By enzyme H ₂ O ₂ Generation of

In the process of the invention H ₂ O ₂ may be produced by an enzyme. It is preferable that the said enzyme is an oxidase. Examples of suitable oxidases include glucose oxidase (EC1.1.3.4), secondary-alcohol oxidase (EC1.1.3.18), methanol oxidase (EC1.1.3.31), oxalate oxidase (EC1.2.3) .4), aryl-aldehyde oxidase (EC1.2.3.9), carbon monoxide oxidase (EC1.2.3.10), amine oxidase (EC1.4.3.4), ethanolamine oxidase (EC1.4.3.8) , Nitroethane oxidase (EC1.7.3.1) and sulfite oxidase (EC1.8.3.1), and the like. Glucose oxidase (EC1.1.3.4) catalyzes the conversion of D-glucose to D-glucono-1,5-lactone and H ₂ O ₂ . Second-alcohol oxidase (EC1.1.3.18) catalyzes the conversion of secondary alcohols to ketones and H ₂ O ₂ . Methanol oxidase (EC1.1.3.31) catalyzes the conversion of methanol to formaldehyde and H ₂ O ₂ . Oxalate oxidase (EC1.2.3.4) catalyzes the conversion of oxalate to carbon dioxide and H ₂ O ₂ . Aryl-aldehyde oxidase (EC1.2.3.9) catalyzes the conversion of aromatic aldehydes to aromatic acids and H ₂ O ₂ . Carbon monoxide oxidase (EC1.2.3.10) catalyzes the conversion of CO and H ₂ O into carbon dioxide and H ₂ O ₂ . Amine oxidase (EC1.4.3.4) catalyzes the conversion of RCH ₂ NH ₂ and H ₂ O to RCHO, NH ₃ and H ₂ O ₂ . Ethanolamine oxidase (EC1.4.3.8) catalyzes the conversion of ethanolamine and H ₂ O to glycolaldehyde and H ₂ O ₂ . Nitroethane oxidase (EC1.7.3.1) catalyzes the conversion of nitroethane and H ₂ O to acetaldehyde and H ₂ O ₂ . Sulfite oxidase (EC1.8.3.1) catalyzes the conversion of sulfite and H ₂ O ₂ to sulfate and H ₂ O ₂ . The oxidase may be purchased commercially available (eg, glucose oxidase). Alternatively, the oxidase may be extracted from known microorganisms according to procedures known in the art.

Substrates of oxidases are known in the art. The H ₂ O ₂ production reaction includes water as well as the substrate. Typically, the reaction also includes a H ₂ O ₂ activating metal. Suitable metals include, but are not limited to, cerium, chromium, cobalt, copper, iron, manganese, molybdenum, silver, titanium, tungsten, vanadium and mixtures thereof. Metallosilicates containing these metals can be prepared and used in the process of the invention. Procedures for making such metallosilicates are known in the art [Neumann et al., Journal of Catalysis, 1997; 166: 206-127. The metallosilicates are preferably titanium in four-sided coordination, for example silicalite-1 (TS-1), silicalite-2 (TS-2), zeolite-beta, ZSM-48 and MCM-41 Silicon analogs of Murugavel and Roesky, Angew. Chem. Int. Ed. Engl., 1997; 36 (5): 477-479].

In a preferred embodiment of the invention, the metal-containing solid or metallosilicate is used as a support to which the H ₂ O ₂ -producing enzyme is immobilized. In another preferred embodiment, the monooxygenase enzyme is also immobilized on the same or different metallosilicate support.

It is preferred to start the reaction by first mixing the oxidase with the other reaction components and then adding the oxidase substrate. For example, the oxidase enzyme is added after mixing all of the monooxygenase enzyme, monooxygenase enzyme substrate and oxidase. In a preferred embodiment, glucose is added after P450 _{BM- 3} , octane and glucose oxidase are mixed together. Typically, control of H ₂ O ₂ production can be performed by controlling the rate of addition of the oxidase substrate.

By precursor H ₂ O ₂ Generation of

In the process of the invention H ₂ O ₂ may be produced by the precursor. It is known in the art to add precursors to water to produce H ₂ O ₂ . Precursors include, but are not limited to, perborate, percarbonate, superphosphate and nitrite peroxide. Preferred precursors are sodium salts. The H ₂ O ₂ -generating properties of the precursors can be enhanced by the use of compounds, for example tetraacetylethylenediamine. The amount of precursor added to the solution containing the monooxygenase enzyme and the substrate is that amount that maximizes the enzymatic reaction with the substrate and minimizes the deactivation of the enzyme by H ₂ O ₂ . The concentration of H ₂ O ₂ produced does not exceed the K _m value of the enzyme, but is sufficient to produce an enzyme reactive species.

Example  One

In this experiment, octane was reacted with electrochemically generated H ₂ O ₂ in the presence of P450 _{BM- 3} heme domain. The experiment was carried out in a 3-electrode shape in a 100 mL glass beaker at room temperature. One vessel contained a reticulated vitreous carbon (RVC) cathode, a platinum gauze anode and an Ag / AgCl reference electrode. The RVC cathode was briefly immersed in a 1 mM 2-aminoanthraquinone ethanol-based solution, taken out and dried in air. The reaction solution contained oxygen saturated aqueous Tris buffer (50 mM, pH 7.4), 0.2 M KCl, 0.5 mM octane and 3 μM P450 _{BM- 3} F87V L188Q A74G heme domain. The reaction solution was stirred to equilibrate (5 to 10 minutes) before -0.55 V vs. A voltage of Ag / AgCl was applied for 2 hours and the solution was continued to stir thoroughly. GC analysis revealed the presence of solvents chloroform, octane, 2-octanol, 3-octanol and 4-octanol and internal standard 1-nonanol. The relative ratio of 2-octanol, 3-octanol and 4-octanol was 1: 1.1: 0.7. The total concentration of octanol formed was 141 μM, indicating a turnover of 47 per enzyme.

Similar experiments were performed using 1.43 μM wild type P450 _{BM- 3} heme domain. The total concentration of octanol formed was 8.4 μM, indicating a turnover of 6 per enzyme. In this case the relative ratio of 2-octanol, 3-octanol and 4-octanol was 1: 1.7: 2.0.

Example  2

In this experiment, octane was reacted with H ₂ O ₂ produced by the enzyme in the presence of P450 _{BM- 3} complete enzyme. The glass vial was filled with a solution consisting of aqueous Tris buffer (50 mM, pH 7.4), 0.5 mM octane, 1.6 μM P450 _{BM- 3} F87V L188Q A74G complete enzyme and glucose oxidase (1.5 U) (total volume 5 mL). After equilibration (5 min), glucose (1 × 10 ⁻⁶ mol) was added to initiate the reaction. Glucose (1 × 10 ⁻⁶ mol) was added successively every 5 minutes up to 1 hour (total 12 additions to 1.2 × 10 ⁻⁵ mol). During this time the reaction was continuously stirred and stopped after 1.5 hours. GC analysis revealed the presence of solvents chloroform, octane, 2-octanol, 3-octanol, 4-octanol and internal standard 1-nonanol. The relative ratio of 2-octanol, 3-octanol and 4-octanol was 1: 1.1: 0.8. The total concentration of octanol formed was 17 μΜ, indicating a turnover of 10 per enzyme.

Example  3

In this experiment, octane was reacted with H ₂ O ₂ derived from sodium perborate in the presence of P450 _{BM- 3} complete enzyme. In a glass vial was added a solution consisting of aqueous Tris buffer (40 mM, pH 7.4), 0.5 mM octane and 1.3 μM P450 _{BM- 3} F87V L188Q A74G complete enzyme (total volume 5 mL). After equilibration (5 _{mins), NaBO 3 .4H 2 O (} 1 × 10 -4 mol) was added and the reaction was initiated, and then stirred for 1 hour. GC analysis revealed the presence of solvents chloroform, octane, 2-octanol, 3-octanol and 4-octanol and internal standard 1-nonanol. The relative ratio of 2-octanol, 3-octanol and 4-octanol was 1: 1.8: 1.1. The total concentration of octanol formed was 77 μΜ, indicating a turnover of 59 per enzyme.

For Examples 1-3, no octanol product was observed when no P450 enzyme was present in the solution.

Example  4

In this experiment, pinene was reacted with H ₂ O ₂ derived from sodium perborate in the presence of a P450 _{BM- 3} heme domain. In a glass vial was placed a solution consisting of aqueous Tris buffer (40 mM, pH 7.4), 0.63 mM pinene and 3.7 μM wild type P450 _{BM- 3} heme domain (5 mL total volume). After equilibration (5 ^{mins), 7.8 × 10 -6 mol NaBO} 3 .4H was added to initiate the reaction the ₂ O, and then stirred for 1 hour. Through GC analysis, cis / trans 2,3-epoxide (32%), (+)-trans-verbenol (16%), (+)-cis-verbenol (6%), (+)-ber The presence of Benon / (+)-Mirthenol (13%), Myrenal (4%) and unidentified additional oxidation product (29%) was found. The total concentration of product formed was 80 μΜ, indicating a turnover of 22 per enzyme.

Example  5

In this experiment, phenol monooxygenase was reacted with phenol in the presence of H ₂ O ₂ produced by sodium perborate. In a glass vial was placed a solution consisting of aqueous Tris buffer (40 mM, pH 7.4), 0.63 mM phenol and 3.7 μM wild type phenol monooxygenase (total volume 5 mL). After equilibration (5 ^{mins), 7.8 × 10 -6 mol NaBO} 3 .4H was added to initiate the reaction the ₂ O, and then stirred for 1 hour. GC analysis revealed the presence of oxidation products.

Example  6

In this experiment, P450 _{BM- 3} was reacted with palmitic acid in the presence of H ₂ O ₂ produced by glucose oxidase. In a glass vial was added a solution consisting of aqueous Tris buffer (50 mM, pH 7.4), 0.5 mM palmitic acid, 1.6 μM P450 _BM-3 complete enzyme and glucose oxidase (1.5 U) (total volume 5 mL). After equilibration (5 min), glucose (1 × 10 ⁻⁶ mol) was added to initiate the reaction. Glucose (1 × 10 ⁻⁶ mol) was added successively every 5 minutes up to 1 hour (total 12 additions to 1.2 × 10 ⁻⁵ mol). During this time the reaction was continuously stirred and stopped after 1.5 hours. GC analysis revealed the presence of oxidation products.

Example  7

Plant CYP74C was reacted with 13 S-hydroperoxylinolenic acid to form compound 3Z-hexenal (aromatic compound). H ₂ O ₂ was produced by sodium perborate. In a glass vial was placed a solution consisting of aqueous Tris buffer (40 mM, pH 7.4), 0.63 mM 13 S-hydroperoxylinolenic acid and 3.7 μΜ wild type plant CYP74C (5 mL total volume). After equilibration (5 ^{mins), 7.8 × 10 -6 mol NaBO} 3 .4H was added to initiate the reaction the ₂ O, and then stirred for 1 hour. GC analysis revealed the presence of oxidation products.

                         SEQUENCE LISTING <110> ISIS INNOVATION LIMITED   <120> HYDROGEN PEROXIDE OXIDATION <130> N.91079A SA <160> 36 <170> PatentIn version 3.2 <210> 1 <211> 1248 <212> DNA (213) Pseudomonas putida <220> <221> CDS (222) (1) .. (1248) <220> <221> misc_feature (222) (4) .. (4) <400> 1 atg acg act gaa acc ata caa agc aac gcc aat ctt gcc cct ctg cca 48 Met Thr Thr Glu Thr Ile Gln Ser Asn Ala Asn Leu Ala Pro Leu Pro 1 5 10 15 ccc cat gtg cca gag cac ctg gta ttc gac ttc gac atg tac aat ccg 96 Pro His Val Pro Glu His Leu Val Phe Asp Phe Asp Met Tyr Asn Pro             20 25 30 tcg aat ctg tct gcc ggc gtg cag gag gcc tgg gca gtt ctg caa gaa 144 Ser Asn Leu Ser Ala Gly Val Gln Glu Ala Trp Ala Val Leu Gln Glu         35 40 45 tca aac gta ccg gat ctg gtg tgg act cgc tgc aac ggc gga cac tgg 192 Ser Asn Val Pro Asp Leu Val Trp Thr Arg Cys Asn Gly Gly His Trp     50 55 60 atc gcc act cgc ggc caa ctg atc cgt gag gcc tat gaa gat tac cgc 240 Ile Ala Thr Arg Gly Gln Leu Ile Arg Glu Ala Tyr Glu Asp Tyr Arg 65 70 75 80 cac ttt tcc agc gag tgc ccg ttc atc cct cgt gaa gcc ggc gaa gcc 288 His Phe Ser Ser Glu Cys Pro Phe Ile Pro Arg Glu Ala Gly Glu Ala                 85 90 95 tac gac ttc att ccc acc tcg atg gat ccg ccc gag cag cgc cag ttt 336 Tyr Asp Phe Ile Pro Thr Ser Met Asp Pro Pro Glu Gln Arg Gln Phe             100 105 110 cgt gcg ctg gcc aac caa gtg gtt ggc atg ccg gtg gtg gat aag ctg 384 Arg Ala Leu Ala Asn Gln Val Val Gly Met Pro Val Val Asp Lys Leu         115 120 125 gag aac cgg atc cag gag ctg gcc tgc tcg ctg atc gag agc ctg cgc 432 Glu Asn Arg Ile Gln Glu Leu Ala Cys Ser Leu Ile Glu Ser Leu Arg     130 135 140 ccg caa gga cag tgc aac ttc acc gag gac tac gcc gaa ccc ttc ccg 480 Pro Gln Gly Gln Cys Asn Phe Thr Glu Asp Tyr Ala Glu Pro Phe Pro 145 150 155 160 ata cgc atc ttc atg ctg ctc gca ggt cta ccg gaa gaa gat atc ccg 528 Ile Arg Ile Phe Met Leu Leu Ala Gly Leu Pro Glu Glu Asp Ile Pro                 165 170 175 cac ttg aaa tac cta acg gat cag atg acc cgt ccg gat ggc agc atg 576 His Leu Lys Tyr Leu Thr Asp Gln Met Thr Arg Pro Asp Gly Ser Met             180 185 190 acc ttc gca gag gcc aag gag gcg ctc tac gac tat ctg ata ccg atc 624 Thr Phe Ala Glu Ala Lys Glu Ala Leu Tyr Asp Tyr Leu Ile Pro Ile         195 200 205 atc gag caa cgc agg cag aag ccg gga acc gac gct atc agc atc gtt 672 Ile Glu Gln Arg Arg Gln Lys Pro Gly Thr Asp Ala Ile Ser Ile Val     210 215 220 gcc aac ggc cag gtc aat ggg cga ccg atc acc agt gac gaa gcc aag 720 Ala Asn Gly Gln Val Asn Gly Arg Pro Ile Thr Ser Asp Glu Ala Lys 225 230 235 240 agg atg tgt ggc ctg tta ctg gtc ggc ggc ctg gat acg gtg gtc aat 768 Arg Met Cys Gly Leu Leu Leu Val Gly Gly Leu Asp Thr Val Val Asn                 245 250 255 ttc ctc agc ttc agc atg gag ttc ctg gcc aaa agc ccg gag cat cgc 816 Phe Leu Ser Phe Ser Met Glu Phe Leu Ala Lys Ser Pro Glu His Arg             260 265 270 cag gag ctg atc gag cgt ccc gag cgt att cca gcc gct tgc gag gaa 864 Gln Glu Leu Ile Glu Arg Pro Glu Arg Ile Pro Ala Ala Cys Glu Glu         275 280 285 cta ctc cgg cgc ttc tcg ctg gtt gcc gat ggc cgc atc ctc acc tcc 912 Leu Leu Arg Arg Phe Ser Leu Val Ala Asp Gly Arg Ile Leu Thr Ser     290 295 300 gat tac gag ttt cat ggc gtg caa ctg aag aaa ggt gac cag atc ctg 960 Asp Tyr Glu Phe His Gly Val Gln Leu Lys Lys Gly Asp Gln Ile Leu 305 310 315 320 cta ccg cag atg ctg tct ggc ctg gat gag cgc gaa aac gcc tgc ccg 1008 Leu Pro Gln Met Leu Ser Gly Leu Asp Glu Arg Glu Asn Ala Cys Pro                 325 330 335 atg cac gtc gac ttc agt cgc caa aag gtt tca cac acc acc ttt ggc 1056 Met His Val Asp Phe Ser Arg Gln Lys Val Ser His Thr Thr Phe Gly             340 345 350 cac ggc agc cat ctg tgc ctt ggc cag cac ctg gcc cgc cgg gaa atc 1104 His Gly Ser His Leu Cys Leu Gly Gln His Leu Ala Arg Arg Glu Ile         355 360 365 atc gtc acc ctc aag gaa tgg ctg acc agg att cct gac ttc tcc att 1152 Ile Val Thr Leu Lys Glu Trp Leu Thr Arg Ile Pro Asp Phe Ser Ile     370 375 380 gcc ccg ggt gcc cag att cag cac aag agc ggc atc gtc agc ggc gtg 1200 Ala Pro Gly Ala Gln Ile Gln His Lys Ser Gly Ile Val Ser Gly Val 385 390 395 400 cag gca ctc cct ctg gtc tgg gat ccg gcg act acc aaa gcg gta taa 1248 Gln Ala Leu Pro Leu Val Trp Asp Pro Ala Thr Thr Lys Ala Val                 405 410 415 <210> 2 <211> 415 <212> PRT (213) Pseudomonas putida <400> 2 Met Thr Thr Glu Thr Ile Gln Ser Asn Ala Asn Leu Ala Pro Leu Pro 1 5 10 15 Pro His Val Pro Glu His Leu Val Phe Asp Phe Asp Met Tyr Asn Pro             20 25 30 Ser Asn Leu Ser Ala Gly Val Gln Glu Ala Trp Ala Val Leu Gln Glu         35 40 45 Ser Asn Val Pro Asp Leu Val Trp Thr Arg Cys Asn Gly Gly His Trp     50 55 60 Ile Ala Thr Arg Gly Gln Leu Ile Arg Glu Ala Tyr Glu Asp Tyr Arg 65 70 75 80 His Phe Ser Ser Glu Cys Pro Phe Ile Pro Arg Glu Ala Gly Glu Ala                 85 90 95 Tyr Asp Phe Ile Pro Thr Ser Met Asp Pro Pro Glu Gln Arg Gln Phe             100 105 110 Arg Ala Leu Ala Asn Gln Val Val Gly Met Pro Val Val Asp Lys Leu         115 120 125 Glu Asn Arg Ile Gln Glu Leu Ala Cys Ser Leu Ile Glu Ser Leu Arg     130 135 140 Pro Gln Gly Gln Cys Asn Phe Thr Glu Asp Tyr Ala Glu Pro Phe Pro 145 150 155 160 Ile Arg Ile Phe Met Leu Leu Ala Gly Leu Pro Glu Glu Asp Ile Pro                 165 170 175 His Leu Lys Tyr Leu Thr Asp Gln Met Thr Arg Pro Asp Gly Ser Met             180 185 190 Thr Phe Ala Glu Ala Lys Glu Ala Leu Tyr Asp Tyr Leu Ile Pro Ile         195 200 205 Ile Glu Gln Arg Arg Gln Lys Pro Gly Thr Asp Ala Ile Ser Ile Val     210 215 220 Ala Asn Gly Gln Val Asn Gly Arg Pro Ile Thr Ser Asp Glu Ala Lys 225 230 235 240 Arg Met Cys Gly Leu Leu Leu Val Gly Gly Leu Asp Thr Val Val Asn                 245 250 255 Phe Leu Ser Phe Ser Met Glu Phe Leu Ala Lys Ser Pro Glu His Arg             260 265 270 Gln Glu Leu Ile Glu Arg Pro Glu Arg Ile Pro Ala Ala Cys Glu Glu         275 280 285 Leu Leu Arg Arg Phe Ser Leu Val Ala Asp Gly Arg Ile Leu Thr Ser     290 295 300 Asp Tyr Glu Phe His Gly Val Gln Leu Lys Lys Gly Asp Gln Ile Leu 305 310 315 320 Leu Pro Gln Met Leu Ser Gly Leu Asp Glu Arg Glu Asn Ala Cys Pro                 325 330 335 Met His Val Asp Phe Ser Arg Gln Lys Val Ser His Thr Thr Phe Gly             340 345 350 His Gly Ser His Leu Cys Leu Gly Gln His Leu Ala Arg Arg Glu Ile         355 360 365 Ile Val Thr Leu Lys Glu Trp Leu Thr Arg Ile Pro Asp Phe Ser Ile     370 375 380 Ala Pro Gly Ala Gln Ile Gln His Lys Ser Gly Ile Val Ser Gly Val 385 390 395 400 Gln Ala Leu Pro Leu Val Trp Asp Pro Ala Thr Thr Lys Ala Val                 405 410 415 <210> 3 <211> 4957 <212> DNA <213> Bacillus megaterium <220> <221> CDS (222) (1541) .. (4690) <400> 3 agatctttat gaagacatag ctgcagaaga aaaagcaaga gctacatatc aatggttaat 60 tgatatatca gatgatcccg atttaaacga cagcttacga tttttacgag aaagagagat 120 tgttcactca cagcggttcc gcgaggccgt ggagatttta aaagatgaca gagacaggaa 180 gaaaatcttt taactagtaa aaaaacatcc cccttggcga atgcaaacga aaggagggat 240 gttttttgtt gtgactgcgt tgattatgcg ctagaactgc agtgacaaga aacaaccttt 300 aatttccctt caacatcttt ccaaactcgc gtataactgt attcacctcc aatagattca 360 ccggttgcca gtgccccatt taacgctact tttgtaacgg taacggcaag ttcttgaaac 420 agtttaactt cttgttccaa cacttccatg cccgctatat caagactttt tgaacgatga 480 acatttatat cttcttcttt tgacaaccat tgcccaaggt gattcacaaa aataagctca 540 tctgaaagta attcttctaa tagctctatg ttattagaaa gcatggctga gcgaagcatt 600 tcttcgtatt ctataactct tgcttgattc atttttaatc ctcctttacg ccttgtgtaa 660 ctcttttcta tttccacgtt gcttttcctt taaacttctt tcattaataa ttcgtgctaa 720 attatgttaa tagaggggat aagtggacta attttctgta agcactaaat attctgaaat 780 actctgttaa ttacctttaa atggtataaa attagaatga aagaaccttt tctttccact 840 tttctagtta tctttttact attaagatgc agttttttat acttgtaatt gtagcggaat 900 gaacgttcat tccgtttttg aaaagaggtg ataaagtgga atctactcca acaaaacaaa 960 aagcgatttt ttctgcttcg cttctgctgt ttgcagaaag agggtttgat gcaaccacga 1020 tgccaatgat tgcagagaat gccaaagtag gagcaggaac aatttatcgc tactttaaaa 1080 ataaagaaag ccttgtaaat gaattattcc aacagcacgt aaacgagttt ttacagtgca 1140 ttgaaagcgg tctggcaaac gagagagatg gataccgaga tgggtttcat catatctttg 1200 aaggtatggt gacatttact aaaaaccatc ctcgtgctct tggatttatt aaaactcata 1260 gccaaggaac ttttttaaca gaagagagcc gcttagcata tcaaaagctg gtggaatttg 1320 tttgtacgtt cttcagagaa ggacaaaagc aaggtgtgat tagaaatctt cctgaaaatg 1380 cgctaattgc tattttattt ggaagtttca tggaagtata tgaaatgatt gaaaatgact 1440 acttatcttt aactgatgaa cttcttaccg gtgtagaaga gagtctgtgg gcagcactta 1500 gcagacaatc atgaaactta acaagtgaaa gagggataac atg aca att aaa gaa 1555                                             Met Thr Ile Lys Glu                                             1 5 atg cct cag cca aaa acg ttt gga gag ctt aaa aat tta ccg tta tta 1603 Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn Leu Pro Leu Leu                 10 15 20 aac aca gat aaa ccg gtt caa gct ttg atg aaa att gcg gat gaa tta 1651 Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile Ala Asp Glu Leu             25 30 35 gga gaa atc ttt aaa ttc gag gcg cct ggt cgt gta acg cgc tac tta 1699 Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val Thr Arg Tyr Leu         40 45 50 tca agt cag cgt cta att aaa gaa gca tgc gat gaa tca cgc ttt gat 1747 Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu Ser Arg Phe Asp     55 60 65 aaa aac tta agt caa gcg ctt aaa ttt gta cgt gat ttt gca gga gac 1795 Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg Asp Phe Ala Gly Asp 70 75 80 85 ggg tta ttt aca agc tgg acg cat gaa aaa aat tgg aaa aaa gcg cat 1843 Gly Leu Phe Thr Ser Trp Thr His Glu Lys Asn Trp Lys Lys Ala His                 90 95 100 aat atc tta ctt cca agc ttc agt cag cag gca atg aaa ggc tat cat 1891 Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met Lys Gly Tyr His             105 110 115 gcg atg atg gtc gat atc gcc gtg cag ctt gtt caa aag tgg gag cgt 1939 Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln Lys Trp Glu Arg         120 125 130 cta aat gca gat gag cat att gaa gta ccg gaa gac atg aca cgt tta 1987 Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp Met Thr Arg Leu     135 140 145 acg ctt gat aca att ggt ctt tgc ggc ttt aac tat cgc ttt aac agc 2035 Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr Arg Phe Asn Ser 150 155 160 165 ttt tac cga gat cag cct cat cca ttt att aca agt atg gtc cgt gca 2083 Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser Met Val Arg Ala                 170 175 180 ctg gat gaa gca atg aac aag ctg cag cga gca aat cca gac gac cca 2131 Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala Asn Pro Asp Asp Pro             185 190 195 gct tat gat gaa aac aag cgc cag ttt caa gaa gat atc aag gtg atg 2179 Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp Ile Lys Val Met         200 205 210 aac gac cta gta gat aaa att att gca gat cgc aaa gca agc ggt gaa 2227 Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys Ala Ser Gly Glu     215 220 225 caa agc gat gat tta tta acg cat atg cta aac gga aaa gat cca gaa 2275 Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly Lys Asp Pro Glu 230 235 240 245 acg ggt gag ccg ctt gat gac gag aac att cgc tat caa att att aca 2323 Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr Gln Ile Ile Thr                 250 255 260 ttc tta att gcg gga cac gaa aca aca agt ggt ctt tta tca ttt gcg 2371 Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu Leu Ser Phe Ala             265 270 275 ctg tat ttc tta gtg aaa aat cca cat gta tta caa aaa gca gca gaa 2419 Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln Lys Ala Ala Glu         280 285 290 gaa gca gca cga gtt cta gta gat cct gtt cca agc tac aaa caa gtc 2467 Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser Tyr Lys Gln Val     295 300 305 aaa cag ctt aaa tat gtc ggc atg gtc tta aac gaa gcg ctg cgc tta 2515 Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu Ala Leu Arg Leu 310 315 320 325 tgg cca act gct cct gcg ttt tcc cta tat gca aaa gaa gat acg gtg 2563 Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys Glu Asp Thr Val                 330 335 340 ctt gga gga gaa tat cct tta gaa aaa ggc gac gaa cta atg gtt ctg 2611 Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu Leu Met Val Leu             345 350 355 att cct cag ctt cac cgt gat aaa aca att tgg gga gac gat gtg gaa 2659 Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly Asp Asp Val Glu         360 365 370 gag ttc cgt cca gag cgt ttt gaa aat cca agt gcg att ccg cag cat 2707 Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala Ile Pro Gln His     375 380 385 gcg ttt aaa ccg ttt gga aac ggt cag cgt gcg tgt atc ggt cag cag 2755 Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys Ile Gly Gln Gln 390 395 400 405 ttc gct ctt cat gaa gca acg ctg gta ctt ggt atg atg cta aaa cac 2803 Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met Met Leu Lys His                 410 415 420 ttt gac ttt gaa gat cat aca aac tac gag ctg gat att aaa gaa act 2851 Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp Ile Lys Glu Thr             425 430 435 tta acg tta aaa cct gaa ggc ttt gtg gta aaa gca aaa tcg aaa aaa 2899 Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala Lys Ser Lys Lys         440 445 450 att ccg ctt ggc ggt att cct tca cct agc act gaa cag tct gct aaa 2947 Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu Gln Ser Ala Lys     455 460 465 aaa gta cgc aaa aag gca gaa aac gct cat aat acg ccg ctg ctt gtg 2995 Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr Pro Leu Leu Val 470 475 480 485 cta tac ggt tca aat atg gga aca gct gaa gga acg gcg cgt gat tta 3043 Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr Ala Arg Asp Leu                 490 495 500 gca gat att gca atg agc aaa gga ttt gca ccg cag gtc gca acg ctt 3091 Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln Val Ala Thr Leu             505 510 515 gat tca cac gcc gga aat ctt ccg cgc gaa gga gct gta tta att gta 3139 Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala Val Leu Ile Val         520 525 530 acg gcg tct tat aac ggt cat ccg cct gat aac gca aag caa ttt gtc 3187 Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala Lys Gln Phe Val     535 540 545 gac tgg tta gac caa gcg tct gct gat gaa gta aaa ggc gtt cgc tac 3235 Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys Gly Val Arg Tyr 550 555 560 565 tcc gta ttt gga tgc ggc gat aaa aac tgg gct act acg tat caa aaa 3283 Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr Thr Tyr Gln Lys                 570 575 580 gtg cct gct ttt atc gat gaa acg ctt gcc gct aaa ggg gca gaa aac 3331 Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys Gly Ala Glu Asn             585 590 595 atc gct gac cgc ggt gaa gca gat gca agc gac gac ttt gaa ggc aca 3379 Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp Phe Glu Gly Thr         600 605 610 tat gaa gaa tgg cgt gaa cat atg tgg agt gac gta gca gcc tac ttt 3427 Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val Ala Ala Tyr Phe     615 620 625 aac ctc gac att gaa aac agt gaa gat aat aaa tct act ctt tca ctt 3475 Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser Thr Leu Ser Leu 630 635 640 645 caa ttt gtc gac agc gcc gcg gat atg ccg ctt gcg aaa atg cac ggt 3523 Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala Lys Met His Gly                 650 655 660 gcg ttt tca acg aac gtc gta gca agc aaa gaa ctt caa cag cca ggc 3571 Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu Gln Gln Pro Gly             665 670 675 agt gca cga agc acg cga cat ctt gaa att gaa ctt cca aaa gaa gct 3619 Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu Pro Lys Glu Ala         680 685 690 tct tat caa gaa gga gat cat tta ggt gtt att cct cgc aac tat gaa 3667 Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro Arg Asn Tyr Glu     695 700 705 gga ata gta aac cgt gta aca gca agg ttc ggc cta gat gca tca cag 3715 Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu Asp Ala Ser Gln 710 715 720 725 caa atc cgt ctg gaa gca gaa gaa gaa aaa tta gct cat ttg cca ctc 3763 Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala His Leu Pro Leu                 730 735 740 gct aaa aca gta tcc gta gaa gag ctt ctg caa tac gtg gag ctt caa 3811 Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr Val Glu Leu Gln             745 750 755 gat cct gtt acg cgc acg cag ctt cgc gca atg gct gct aaa acg gtc 3859 Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala Ala Lys Thr Val         760 765 770 tgc ccg ccg cat aaa gta gag ctt gaa gcc ttg ctt gaa aag caa gcc 3907 Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu Glu Lys Gln Ala     775 780 785 tac aaa gaa caa gtg ctg gca aaa cgt tta aca atg ctt gaa ctg ctt 3955 Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met Leu Glu Leu Leu 790 795 800 805 gaa aaa tac ccg gcg tgt gaa atg aaa ttc agc gaa ttt atc gcc ctt 4003 Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu Phe Ile Ala Leu                 810 815 820 ctg cca agc ata cgc ccg cgc tat tac tcg att tct tca tca cct cgt 4051 Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser Ser Ser Pro Arg             825 830 835 gtc gat gaa aaa caa gca agc atc acg gtc agc gtt gtc tca gga gaa 4099 Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val Val Ser Gly Glu         840 845 850 gcg tgg agc gga tat gga gaa tat aaa gga att gcg tcg aac tat ctt 4147 Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala Ser Asn Tyr Leu     855 860 865 gcc gag ctg caa gaa gga gat acg att acg tgc ttt att tcc aca ccg 4195 Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe Ile Ser Thr Pro 870 875 880 885 cag tca gaa ttt acg ctg cca aaa gac cct gaa acg ccg ctt atc atg 4243 Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr Pro Leu Ile Met                 890 895 900 gtc gga ccg gga aca ggc gtc gcg ccg ttt aga ggc ttt gtg cag gcg 4291 Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly Phe Val Gln Ala             905 910 915 cgc aaa cag cta aaa gaa caa gga cag tca ctt gga gaa gca cat tta 4339 Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly Glu Ala His Leu         920 925 930 tac ttc ggc tgc cgt tca cct cat gaa gac tat ctg tat caa gaa gag 4387 Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu Tyr Gln Glu Glu     935 940 945 ctt gaa aac gcc caa agc gaa ggc atc att acg ctt cat acc gct ttt 4435 Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu His Thr Ala Phe 950 955 960 965 tct cgc atg cca aat cag ccg aaa aca tac gtt cag cac gta atg gaa 4483 Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln His Val Met Glu                 970 975 980 caa gac ggc aag aaa ttg att gaa ctt ctt gat caa gga gcg cac ttc 4531 Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln Gly Ala His Phe             985 990 995 tat att tgc gga gac gga agc caa atg gca cct gcc gtt gaa gca 4576 Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala Val Glu Ala         1000 1005 1010 acg ctt atg aaa agc tat gct gac gtt cac caa gtg agt gaa gca 4621 Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val Ser Glu Ala         1015 1020 1025 gac gct cgc tta tgg ctg cag cag cta gaa gaa aaa ggc cga tac 4666 Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys Gly Arg Tyr         1030 1035 1040 gca aaa gac gtg tgg gct ggg taa attaaaaaga ggctaggata aaagtagttt 4720 Ala Lys Asp Val Trp Ala Gly         1045 agttggttga aggaagatcc gaacgatgaa tcgttcggat ctttttattg gtagagtaaa 4780 cgtagatttc atctatttag tgacttgtag cggttgattg gagggcaagg tgaagactcc 4840 aatcaaccgc ggtgtcacat gcaagccata cgaaattcat ttctcccatt tattcgtctt 4900 ttgtccccac ttaattttta tagcgcctta acgtttcttc tgcgtgacag cagatct 4957 <210> 4 <211> 1049 <212> PRT <213> Bacillus megaterium <400> 4 Met Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys 1 5 10 15 Asn Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys             20 25 30 Ile Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg         35 40 45 Val Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp     50 55 60 Glu Ser Arg Phe Asp Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg 65 70 75 80 Asp Phe Ala Gly Asp Gly Leu Phe Thr Ser Trp Thr His Glu Lys Asn                 85 90 95 Trp Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala             100 105 110 Met Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val         115 120 125 Gln Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu     130 135 140 Asp Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn 145 150 155 160 Tyr Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr                 165 170 175 Ser Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala             180 185 190 Asn Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu         195 200 205 Asp Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg     210 215 220 Lys Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn 225 230 235 240 Gly Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg                 245 250 255 Tyr Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly             260 265 270 Leu Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu         275 280 285 Gln Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro     290 295 300 Ser Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn 305 310 315 320 Glu Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala                 325 330 335 Lys Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp             340 345 350 Glu Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp         355 360 365 Gly Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser     370 375 380 Ala Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala 385 390 395 400 Cys Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly                 405 410 415 Met Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu             420 425 430 Asp Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys         435 440 445 Ala Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr     450 455 460 Glu Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn 465 470 475 480 Thr Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly                 485 490 495 Thr Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro             500 505 510 Gln Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly         515 520 525 Ala Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn     530 535 540 Ala Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val 545 550 555 560 Lys Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala                 565 570 575 Thr Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala             580 585 590 Lys Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp         595 600 605 Asp Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp     610 615 620 Val Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys 625 630 635 640 Ser Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu                 645 650 655 Ala Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu             660 665 670 Leu Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu         675 680 685 Leu Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile     690 695 700 Pro Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly 705 710 715 720 Leu Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu                 725 730 735 Ala His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln             740 745 750 Tyr Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met         755 760 765 Ala Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu     770 775 780 Leu Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr 785 790 795 800 Met Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser                 805 810 815 Glu Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile             820 825 830 Ser Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser         835 840 845 Val Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile     850 855 860 Ala Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys 865 870 875 880 Phe Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu                 885 890 895 Thr Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg             900 905 910 Gly Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu         915 920 925 Gly Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr     930 935 940 Leu Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr 945 950 955 960 Leu His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val                 965 970 975 Gln His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp             980 985 990 Gln Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro         995 1000 1005 Ala Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln     1010 1015 1020 Val Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu     1025 1030 1035 Lys Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly     1040 1045 <210> 5 <211> 3147 <212> DNA <213> Artificial sequence <220> <223> Cytochrome P450BM-3 mutant <220> <221> CDS (222) (1) .. (3147) <400> 5 aca att aaa gaa atg cct cag cca aaa acg ttt gga gag ctt aaa aat 48 Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn 1 5 10 15 tta ccg tta tta aac aca gat aaa ccg gtt caa gct ttg atg aaa att 96 Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile             20 25 30 gcg gat gaa tta gga gaa atc ttt aaa ttc gag gcg cct ggt cgt gta 144 Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val         35 40 45 acg cgc tac tta tca agt cag cgt cta att aaa gaa gca tgc gat gaa 192 Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu     50 55 60 tca cgc ttt gat aaa aac tta agt caa gcg ctt aaa ttt gta cgt gat 240 Ser Arg Phe Asp Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg Asp 65 70 75 80 ttt gca gga gac ggg tta gct aca agc tgg acg cat gaa aaa aat tgg 288 Phe Ala Gly Asp Gly Leu Ala Thr Ser Trp Thr His Glu Lys Asn Trp                 85 90 95 aaa aaa gcg cat aat atc tta ctt cca agc ttc agt cag cag gca atg 336 Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met             100 105 110 aaa ggc tat cat gcg atg atg gtc gat atc gcc gtg cag ctt gtt caa 384 Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln         115 120 125 aag tgg gag cgt cta aat gca gat gag cat att gaa gta ccg gaa gac 432 Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp     130 135 140 atg aca cgt tta acg ctt gat aca att ggt ctt tgc ggc ttt aac tat 480 Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr 145 150 155 160 cgc ttt aac agc ttt tac cga gat cag cct cat cca ttt att aca agt 528 Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser                 165 170 175 atg gtc cgt gca ctg gat gaa gca atg aac aag ctg cag cga gca aat 576 Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala Asn             180 185 190 cca gac gac cca gct tat gat gaa aac aag cgc cag ttt caa gaa gat 624 Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp         195 200 205 atc aag gtg atg aac gac cta gta gat aaa att att gca gat cgc aaa 672 Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys     210 215 220 gca agc ggt gaa caa agc gat gat tta tta acg cat atg cta aac gga 720 Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly 225 230 235 240 aaa gat cca gaa acg ggt gag ccg ctt gat gac gag aac att cgc tat 768 Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr                 245 250 255 caa att att aca ttc tta att gcg gga cac gaa aca aca agt ggt ctt 816 Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu             260 265 270 tta tca ttt gcg ctg tat ttc tta gtg aaa aat cca cat gta tta caa 864 Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln         275 280 285 aaa gca gca gaa gaa gca gca cga gtt cta gta gat cct gtt cca agc 912 Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser     290 295 300 tac aaa caa gtc aaa cag ctt aaa tat gtc ggc atg gtc tta aac gaa 960 Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu 305 310 315 320 gcg ctg cgc tta tgg cca act gct cct gcg ttt tcc cta tat gca aaa 1008 Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys                 325 330 335 gaa gat acg gtg ctt gga gga gaa tat cct tta gaa aaa ggc gac gaa 1056 Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu             340 345 350 cta atg gtt ctg att cct cag ctt cac cgt gat aaa aca att tgg gga 1104 Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly         355 360 365 gac gat gtg gaa gag ttc cgt cca gag cgt ttt gaa aat cca agt gcg 1152 Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala     370 375 380 att ccg cag cat gcg ttt aaa ccg ttt gga aac ggt cag cgt gcg tgt 1200 Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys 385 390 395 400 atc ggt cag cag ttc gct ctt cat gaa gca acg ctg gta ctt ggt atg 1248 Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met                 405 410 415 atg cta aaa cac ttt gac ttt gaa gat cat aca aac tac gag ctg gat 1296 Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp             420 425 430 att aaa gaa act tta acg tta aaa cct gaa ggc ttt gtg gta aaa gca 1344 Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala         435 440 445 aaa tcg aaa aaa att ccg ctt ggc ggt att cct tca cct agc act gaa 1392 Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu     450 455 460 cag tct gct aaa aaa gta cgc aaa aag gca gaa aac gct cat aat acg 1440 Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr 465 470 475 480 ccg ctg ctt gtg cta tac ggt tca aat atg gga aca gct gaa gga acg 1488 Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr                 485 490 495 gcg cgt gat tta gca gat att gca atg agc aaa gga ttt gca ccg cag 1536 Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln             500 505 510 gtc gca acg ctt gat tca cac gcc gga aat ctt ccg cgc gaa gga gct 1584 Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala         515 520 525 gta tta att gta acg gcg tct tat aac ggt cat ccg cct gat aac gca 1632 Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala     530 535 540 aag caa ttt gtc gac tgg tta gac caa gcg tct gct gat gaa gta aaa 1680 Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys 545 550 555 560 ggc gtt cgc tac tcc gta ttt gga tgc ggc gat aaa aac tgg gct act 1728 Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr                 565 570 575 acg tat caa aaa gtg cct gct ttt atc gat gaa acg ctt gcc gct aaa 1776 Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys             580 585 590 ggg gca gaa aac atc gct gac cgc ggt gaa gca gat gca agc gac gac 1824 Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp         595 600 605 ttt gaa ggc aca tat gaa gaa tgg cgt gaa cat atg tgg agt gac gta 1872 Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val     610 615 620 gca gcc tac ttt aac ctc gac att gaa aac agt gaa gat aat aaa tct 1920 Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser 625 630 635 640 act ctt tca ctt caa ttt gtc gac agc gcc gcg gat atg ccg ctt gcg 1968 Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala                 645 650 655 aaa atg cac ggt gcg ttt tca acg aac gtc gta gca agc aaa gaa ctt 2016 Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu             660 665 670 caa cag cca ggc agt gca cga agc acg cga cat ctt gaa att gaa ctt 2064 Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu         675 680 685 cca aaa gaa gct tct tat caa gaa gga gat cat tta ggt gtt att cct 2112 Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro     690 695 700 cgc aac tat gaa gga ata gta aac cgt gta aca gca agg ttc ggc cta 2160 Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu 705 710 715 720 gat gca tca cag caa atc cgt ctg gaa gca gaa gaa gaa aaa tta gct 2208 Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala                 725 730 735 cat ttg cca ctc gct aaa aca gta tcc gta gaa gag ctt ctg caa tac 2256 His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr             740 745 750 gtg gag ctt caa gat cct gtt acg cgc acg cag ctt cgc gca atg gct 2304 Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala         755 760 765 gct aaa acg gtc tgc ccg ccg cat aaa gta gag ctt gaa gcc ttg ctt 2352 Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu     770 775 780 gaa aag caa gcc tac aaa gaa caa gtg ctg gca aaa cgt tta aca atg 2400 Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met 785 790 795 800 ctt gaa ctg ctt gaa aaa tac ccg gcg tgt gaa atg aaa ttc agc gaa 2448 Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu                 805 810 815 ttt atc gcc ctt ctg cca agc ata cgc ccg cgc tat tac tcg att tct 2496 Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser             820 825 830 tca tca cct cgt gtc gat gaa aaa caa gca agc atc acg gtc agc gtt 2544 Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val         835 840 845 gtc tca gga gaa gcg tgg agc gga tat gga gaa tat aaa gga att gcg 2592 Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala     850 855 860 tcg aac tat ctt gcc gag ctg caa gaa gga gat acg att acg tgc ttt 2640 Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe 865 870 875 880 att tcc aca ccg cag tca gaa ttt acg ctg cca aaa gac cct gaa acg 2688 Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr                 885 890 895 ccg ctt atc atg gtc gga ccg gga aca ggc gtc gcg ccg ttt aga ggc 2736 Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly             900 905 910 ttt gtg cag gcg cgc aaa cag cta aaa gaa caa gga cag tca ctt gga 2784 Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly         915 920 925 gaa gca cat tta tac ttc ggc tgc cgt tca cct cat gaa gac tat ctg 2832 Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu     930 935 940 tat caa gaa gag ctt gaa aac gcc caa agc gaa ggc atc att acg ctt 2880 Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu 945 950 955 960 cat acc gct ttt tct cgc atg cca aat cag ccg aaa aca tac gtt cag 2928 His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln                 965 970 975 cac gta atg gaa caa gac ggc aag aaa ttg att gaa ctt ctt gat caa 2976 His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln             980 985 990 gga gcg cac ttc tat att tgc gga gac gga agc caa atg gca cct gcc 3024 Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala         995 1000 1005 gtt gaa gca acg ctt atg aaa agc tat gct gac gtt cac caa gtg 3069 Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val     1010 1015 1020 agt gaa gca gac gct cgc tta tgg ctg cag cag cta gaa gaa aaa 3114 Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys     1025 1030 1035 ggc cga tac gca aaa gac gtg tgg gct ggg taa 3147 Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly     1040 1045 <210> 6 <211> 1048 <212> PRT <213> Artificial sequence <220> <223> Synthetic Construct <400> 6 Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn 1 5 10 15 Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile             20 25 30 Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val         35 40 45 Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu     50 55 60 Ser Arg Phe Asp Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg Asp 65 70 75 80 Phe Ala Gly Asp Gly Leu Ala Thr Ser Trp Thr His Glu Lys Asn Trp                 85 90 95 Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met             100 105 110 Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln         115 120 125 Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp     130 135 140 Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr 145 150 155 160 Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser                 165 170 175 Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala Asn             180 185 190 Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp         195 200 205 Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys     210 215 220 Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly 225 230 235 240 Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr                 245 250 255 Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu             260 265 270 Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln         275 280 285 Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser     290 295 300 Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu 305 310 315 320 Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys                 325 330 335 Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu             340 345 350 Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly         355 360 365 Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala     370 375 380 Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys 385 390 395 400 Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met                 405 410 415 Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp             420 425 430 Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala         435 440 445 Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu     450 455 460 Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr 465 470 475 480 Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr                 485 490 495 Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln             500 505 510 Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala         515 520 525 Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala     530 535 540 Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys 545 550 555 560 Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr                 565 570 575 Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys             580 585 590 Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp         595 600 605 Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val     610 615 620 Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser 625 630 635 640 Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala                 645 650 655 Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu             660 665 670 Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu         675 680 685 Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro     690 695 700 Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu 705 710 715 720 Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala                 725 730 735 His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr             740 745 750 Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala         755 760 765 Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu     770 775 780 Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met 785 790 795 800 Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu                 805 810 815 Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser             820 825 830 Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val         835 840 845 Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala     850 855 860 Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe 865 870 875 880 Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr                 885 890 895 Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly             900 905 910 Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly         915 920 925 Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu     930 935 940 Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu 945 950 955 960 His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln                 965 970 975 His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln             980 985 990 Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala         995 1000 1005 Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val     1010 1015 1020 Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys     1025 1030 1035 Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly     1040 1045 <210> 7 <211> 3147 <212> DNA <213> Artificial sequence <220> <223> Cytochrome P450BM-3 mutant <220> <221> CDS (222) (1) .. (3147) <400> 7 aca att aaa gaa atg cct cag cca aaa acg ttt gga gag ctt aaa aat 48 Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn 1 5 10 15 tta ccg tta tta aac aca gat aaa ccg gtt caa gct ttg atg aaa att 96 Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile             20 25 30 gcg gat gaa tta gga gaa atc ttt aaa ttc gag gcg cct ggt cgt gta 144 Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val         35 40 45 acg cgc tac tta tca agt cag cgt cta att aaa gaa gca tgc gat gaa 192 Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu     50 55 60 tca cgc ttt gat aaa aac tta agt caa ggg ctt aaa ttt gta cgt gat 240 Ser Arg Phe Asp Lys Asn Leu Ser Gln Gly Leu Lys Phe Val Arg Asp 65 70 75 80 ttt gca gga gac ggg tta gtt aca agc tgg acg cat gaa aaa aat tgg 288 Phe Ala Gly Asp Gly Leu Val Thr Ser Trp Thr His Glu Lys Asn Trp                 85 90 95 aaa aaa gcg cat aat atc tta ctt cca agc ttc agt cag cag gca atg 336 Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met             100 105 110 aaa ggc tat cat gcg atg atg gtc gat atc gcc gtg cag ctt gtt caa 384 Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln         115 120 125 aag tgg gag cgt cta aat gca gat gag cat att gaa gta ccg gaa gac 432 Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp     130 135 140 atg aca cgt tta acg ctt gat aca att ggt ctt tgc ggc ttt aac tat 480 Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr 145 150 155 160 cgc ttt aac agc ttt tac cga gat cag cct cat cca ttt att aca agt 528 Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser                 165 170 175 atg gtc cgt gca ctg gat gaa gca atg aac aag cag cag cga gca aat 576 Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Gln Gln Arg Ala Asn             180 185 190 cca gac gac cca gct tat gat gaa aac aag cgc cag ttt caa gaa gat 624 Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp         195 200 205 atc aag gtg atg aac gac cta gta gat aaa att att gca gat cgc aaa 672 Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys     210 215 220 gca agc ggt gaa caa agc gat gat tta tta acg cat atg cta aac gga 720 Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly 225 230 235 240 aaa gat cca gaa acg ggt gag ccg ctt gat gac gag aac att cgc tat 768 Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr                 245 250 255 caa att att aca ttc tta att gcg gga cac gaa aca aca agt ggt ctt 816 Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu             260 265 270 tta tca ttt gcg ctg tat ttc tta gtg aaa aat cca cat gta tta caa 864 Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln         275 280 285 aaa gca gca gaa gaa gca gca cga gtt cta gta gat cct gtt cca agc 912 Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser     290 295 300 tac aaa caa gtc aaa cag ctt aaa tat gtc ggc atg gtc tta aac gaa 960 Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu 305 310 315 320 gcg ctg cgc tta tgg cca act gct cct gcg ttt tcc cta tat gca aaa 1008 Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys                 325 330 335 gaa gat acg gtg ctt gga gga gaa tat cct tta gaa aaa ggc gac gaa 1056 Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu             340 345 350 cta atg gtt ctg att cct cag ctt cac cgt gat aaa aca att tgg gga 1104 Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly         355 360 365 gac gat gtg gaa gag ttc cgt cca gag cgt ttt gaa aat cca agt gcg 1152 Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala     370 375 380 att ccg cag cat gcg ttt aaa ccg ttt gga aac ggt cag cgt gcg tgt 1200 Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys 385 390 395 400 atc ggt cag cag ttc gct ctt cat gaa gca acg ctg gta ctt ggt atg 1248 Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met                 405 410 415 atg cta aaa cac ttt gac ttt gaa gat cat aca aac tac gag ctg gat 1296 Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp             420 425 430 att aaa gaa act tta acg tta aaa cct gaa ggc ttt gtg gta aaa gca 1344 Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala         435 440 445 aaa tcg aaa aaa att ccg ctt ggc ggt att cct tca cct agc act gaa 1392 Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu     450 455 460 cag tct gct aaa aaa gta cgc aaa aag gca gaa aac gct cat aat acg 1440 Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr 465 470 475 480 ccg ctg ctt gtg cta tac ggt tca aat atg gga aca gct gaa gga acg 1488 Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr                 485 490 495 gcg cgt gat tta gca gat att gca atg agc aaa gga ttt gca ccg cag 1536 Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln             500 505 510 gtc gca acg ctt gat tca cac gcc gga aat ctt ccg cgc gaa gga gct 1584 Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala         515 520 525 gta tta att gta acg gcg tct tat aac ggt cat ccg cct gat aac gca 1632 Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala     530 535 540 aag caa ttt gtc gac tgg tta gac caa gcg tct gct gat gaa gta aaa 1680 Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys 545 550 555 560 ggc gtt cgc tac tcc gta ttt gga tgc ggc gat aaa aac tgg gct act 1728 Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr                 565 570 575 acg tat caa aaa gtg cct gct ttt atc gat gaa acg ctt gcc gct aaa 1776 Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys             580 585 590 ggg gca gaa aac atc gct gac cgc ggt gaa gca gat gca agc gac gac 1824 Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp         595 600 605 ttt gaa ggc aca tat gaa gaa tgg cgt gaa cat atg tgg agt gac gta 1872 Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val     610 615 620 gca gcc tac ttt aac ctc gac att gaa aac agt gaa gat aat aaa tct 1920 Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser 625 630 635 640 act ctt tca ctt caa ttt gtc gac agc gcc gcg gat atg ccg ctt gcg 1968 Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala                 645 650 655 aaa atg cac ggt gcg ttt tca acg aac gtc gta gca agc aaa gaa ctt 2016 Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu             660 665 670 caa cag cca ggc agt gca cga agc acg cga cat ctt gaa att gaa ctt 2064 Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu         675 680 685 cca aaa gaa gct tct tat caa gaa gga gat cat tta ggt gtt att cct 2112 Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro     690 695 700 cgc aac tat gaa gga ata gta aac cgt gta aca gca agg ttc ggc cta 2160 Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu 705 710 715 720 gat gca tca cag caa atc cgt ctg gaa gca gaa gaa gaa aaa tta gct 2208 Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala                 725 730 735 cat ttg cca ctc gct aaa aca gta tcc gta gaa gag ctt ctg caa tac 2256 His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr             740 745 750 gtg gag ctt caa gat cct gtt acg cgc acg cag ctt cgc gca atg gct 2304 Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala         755 760 765 gct aaa acg gtc tgc ccg ccg cat aaa gta gag ctt gaa gcc ttg ctt 2352 Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu     770 775 780 gaa aag caa gcc tac aaa gaa caa gtg ctg gca aaa cgt tta aca atg 2400 Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met 785 790 795 800 ctt gaa ctg ctt gaa aaa tac ccg gcg tgt gaa atg aaa ttc agc gaa 2448 Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu                 805 810 815 ttt atc gcc ctt ctg cca agc ata cgc ccg cgc tat tac tcg att tct 2496 Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser             820 825 830 tca tca cct cgt gtc gat gaa aaa caa gca agc atc acg gtc agc gtt 2544 Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val         835 840 845 gtc tca gga gaa gcg tgg agc gga tat gga gaa tat aaa gga att gcg 2592 Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala     850 855 860 tcg aac tat ctt gcc gag ctg caa gaa gga gat acg att acg tgc ttt 2640 Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe 865 870 875 880 att tcc aca ccg cag tca gaa ttt acg ctg cca aaa gac cct gaa acg 2688 Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr                 885 890 895 ccg ctt atc atg gtc gga ccg gga aca ggc gtc gcg ccg ttt aga ggc 2736 Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly             900 905 910 ttt gtg cag gcg cgc aaa cag cta aaa gaa caa gga cag tca ctt gga 2784 Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly         915 920 925 gaa gca cat tta tac ttc ggc tgc cgt tca cct cat gaa gac tat ctg 2832 Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu     930 935 940 tat caa gaa gag ctt gaa aac gcc caa agc gaa ggc atc att acg ctt 2880 Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu 945 950 955 960 cat acc gct ttt tct cgc atg cca aat cag ccg aaa aca tac gtt cag 2928 His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln                 965 970 975 cac gta atg gaa caa gac ggc aag aaa ttg att gaa ctt ctt gat caa 2976 His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln             980 985 990 gga gcg cac ttc tat att tgc gga gac gga agc caa atg gca cct gcc 3024 Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala         995 1000 1005 gtt gaa gca acg ctt atg aaa agc tat gct gac gtt cac caa gtg 3069 Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val     1010 1015 1020 agt gaa gca gac gct cgc tta tgg ctg cag cag cta gaa gaa aaa 3114 Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys     1025 1030 1035 ggc cga tac gca aaa gac gtg tgg gct ggg taa 3147 Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly     1040 1045 <210> 8 <211> 1048 <212> PRT <213> Artificial sequence <220> <223> Synthetic Construct <400> 8 Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn 1 5 10 15 Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile             20 25 30 Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val         35 40 45 Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu     50 55 60 Ser Arg Phe Asp Lys Asn Leu Ser Gln Gly Leu Lys Phe Val Arg Asp 65 70 75 80 Phe Ala Gly Asp Gly Leu Val Thr Ser Trp Thr His Glu Lys Asn Trp                 85 90 95 Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met             100 105 110 Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln         115 120 125 Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp     130 135 140 Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr 145 150 155 160 Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser                 165 170 175 Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Gln Gln Arg Ala Asn             180 185 190 Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp         195 200 205 Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys     210 215 220 Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly 225 230 235 240 Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr                 245 250 255 Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu             260 265 270 Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln         275 280 285 Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser     290 295 300 Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu 305 310 315 320 Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys                 325 330 335 Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu             340 345 350 Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly         355 360 365 Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala     370 375 380 Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys 385 390 395 400 Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met                 405 410 415 Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp             420 425 430 Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala         435 440 445 Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu     450 455 460 Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr 465 470 475 480 Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr                 485 490 495 Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln             500 505 510 Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala         515 520 525 Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala     530 535 540 Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys 545 550 555 560 Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr                 565 570 575 Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys             580 585 590 Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp         595 600 605 Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val     610 615 620 Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser 625 630 635 640 Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala                 645 650 655 Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu             660 665 670 Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu         675 680 685 Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro     690 695 700 Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu 705 710 715 720 Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala                 725 730 735 His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr             740 745 750 Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala         755 760 765 Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu     770 775 780 Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met 785 790 795 800 Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu                 805 810 815 Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser             820 825 830 Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val         835 840 845 Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala     850 855 860 Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe 865 870 875 880 Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr                 885 890 895 Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly             900 905 910 Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly         915 920 925 Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu     930 935 940 Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu 945 950 955 960 His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln                 965 970 975 His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln             980 985 990 Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala         995 1000 1005 Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val     1010 1015 1020 Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys     1025 1030 1035 Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly     1040 1045 <210> 9 <211> 1032 <212> DNA <213> Nocardia corallina <220> <221> CDS (222) (1) .. (1032) <400> 9 atg acg aca gag gcg acg gtg gcc cga ccg gtg gag ctc gaa ggt cac 48 Met Thr Thr Glu Ala Thr Val Ala Arg Pro Val Glu Leu Glu Gly His 1 5 10 15 cgg aca ttc acc tgg ttc acg ccc gcc agg cga aag ccg acg gag tac 96 Arg Thr Phe Thr Trp Phe Thr Pro Ala Arg Arg Lys Pro Thr Glu Tyr             20 25 30 gag ctc tac acc gtg ggt caa cag tcc act ccg gac gag tgg ctg cat 144 Glu Leu Tyr Thr Val Gly Gln Gln Ser Thr Pro Asp Glu Trp Leu His         35 40 45 gtg gac tgg ccg ctg cgc ttc gac gac ggc cgc gcc ccg tgg gag gag 192 Val Asp Trp Pro Leu Arg Phe Asp Asp Gly Arg Ala Pro Trp Glu Glu     50 55 60 gag tcg agt gcg gta cgg acc tcg gag tgg tcg gct tac cgc gac cca 240 Glu Ser Ser Ala Val Arg Thr Ser Glu Trp Ser Ala Tyr Arg Asp Pro 65 70 75 80 cac caa ctg tgg cag cgt ccc tac gtc agc acg tgc aac cag gac cag 288 His Gln Leu Trp Gln Arg Pro Tyr Val Ser Thr Cys Asn Gln Asp Gln                 85 90 95 cag gcc ctc gcg cgg ctg gtc ccc gtc ctg acc atg ggg tcg gcg gcg 336 Gln Ala Leu Ala Arg Leu Val Pro Val Leu Thr Met Gly Ser Ala Ala             100 105 110 atc acg ccc atc tgg tcg cag aag atc ctc gcc agg tcc tac gcc gcc 384 Ile Thr Pro Ile Trp Ser Gln Lys Ile Leu Ala Arg Ser Tyr Ala Ala         115 120 125 tgg cca ttc gtc gag tac ggg ctc ttc ctg agc ctg gcc tac gcc gtg 432 Trp Pro Phe Val Glu Tyr Gly Leu Phe Leu Ser Leu Ala Tyr Ala Val     130 135 140 cgc cag gcc atg tcc gac acg gtc cag ttc agc gtg gtg ttc cag gcc 480 Arg Gln Ala Met Ser Asp Thr Val Gln Phe Ser Val Val Phe Gln Ala 145 150 155 160 gtg gac cgc atg cgg ctg ctc cag gac atc gtc cac cac ctg gac cac 528 Val Asp Arg Met Arg Leu Leu Gln Asp Ile Val His His Leu Asp His                 165 170 175 ctg cag gag tcg ccg gaa ttc agc gac gcc ggg gcc cgc gag gcc tgg 576 Leu Gln Glu Ser Pro Glu Phe Ser Asp Ala Gly Ala Arg Glu Ala Trp             180 185 190 atg tcc gac tcc acc ctg gtc ccg atc cgg gaa gtg atc gag cgc atc 624 Met Ser Asp Ser Thr Leu Val Pro Ile Arg Glu Val Ile Glu Arg Ile         195 200 205 gcc gcc agc cag gac tgg gtg gag atc ctg gtc gcc ggc acg ctc gtc 672 Ala Ala Ser Gln Asp Trp Val Glu Ile Leu Val Ala Gly Thr Leu Val     210 215 220 ttc gag cct ctg gtc ggc cac ctg gcg aag gcc gag ttg ttc agc cgc 720 Phe Glu Pro Leu Val Gly His Leu Ala Lys Ala Glu Leu Phe Ser Arg 225 230 235 240 cgt gcg cca atg ttc ggg gac ggg acc acg ccg gcg gtg ctg gcg tcg 768 Arg Ala Pro Met Phe Gly Asp Gly Thr Thr Pro Ala Val Leu Ala Ser                 245 250 255 gcc ctg ctg gac agc ggc agg cac ctc gaa tcg gtc cag gcg ctc gtc 816 Ala Leu Leu Asp Ser Gly Arg His Leu Glu Ser Val Gln Ala Leu Val             260 265 270 cgc ctc gtc tgc caa gac ccc gtc cat ggc gac cag aac cag gcg act 864 Arg Leu Val Cys Gln Asp Pro Val His Gly Asp Gln Asn Gln Ala Thr         275 280 285 gtg cgg cgg tgg atc gag gaa tgg cag ccg cgg tgc aag gcg gcg gcc 912 Val Arg Arg Trp Ile Glu Glu Trp Gln Pro Arg Cys Lys Ala Ala Ala     290 295 300 cag tcc ttc ctg ccg acg ttc tcc gac tgc ggc atc gac gcc aag gaa 960 Gln Ser Phe Leu Pro Thr Phe Ser Asp Cys Gly Ile Asp Ala Lys Glu 305 310 315 320 agc gcc aac gcg ctg tcc cgg gcg ctg gcg aac cag cgg gcc gcc gtc 1008 Ser Ala Asn Ala Leu Ser Arg Ala Leu Ala Asn Gln Arg Ala Ala Val                 325 330 335 gag ggc gcc ggc atc acg gca tga 1032 Glu Gly Ala Gly Ile Thr Ala             340 <210> 10 <211> 343 <212> PRT <213> Nocardia corallina <400> 10 Met Thr Thr Glu Ala Thr Val Ala Arg Pro Val Glu Leu Glu Gly His 1 5 10 15 Arg Thr Phe Thr Trp Phe Thr Pro Ala Arg Arg Lys Pro Thr Glu Tyr             20 25 30 Glu Leu Tyr Thr Val Gly Gln Gln Ser Thr Pro Asp Glu Trp Leu His         35 40 45 Val Asp Trp Pro Leu Arg Phe Asp Asp Gly Arg Ala Pro Trp Glu Glu     50 55 60 Glu Ser Ser Ala Val Arg Thr Ser Glu Trp Ser Ala Tyr Arg Asp Pro 65 70 75 80 His Gln Leu Trp Gln Arg Pro Tyr Val Ser Thr Cys Asn Gln Asp Gln                 85 90 95 Gln Ala Leu Ala Arg Leu Val Pro Val Leu Thr Met Gly Ser Ala Ala             100 105 110 Ile Thr Pro Ile Trp Ser Gln Lys Ile Leu Ala Arg Ser Tyr Ala Ala         115 120 125 Trp Pro Phe Val Glu Tyr Gly Leu Phe Leu Ser Leu Ala Tyr Ala Val     130 135 140 Arg Gln Ala Met Ser Asp Thr Val Gln Phe Ser Val Val Phe Gln Ala 145 150 155 160 Val Asp Arg Met Arg Leu Leu Gln Asp Ile Val His His Leu Asp His                 165 170 175 Leu Gln Glu Ser Pro Glu Phe Ser Asp Ala Gly Ala Arg Glu Ala Trp             180 185 190 Met Ser Asp Ser Thr Leu Val Pro Ile Arg Glu Val Ile Glu Arg Ile         195 200 205 Ala Ala Ser Gln Asp Trp Val Glu Ile Leu Val Ala Gly Thr Leu Val     210 215 220 Phe Glu Pro Leu Val Gly His Leu Ala Lys Ala Glu Leu Phe Ser Arg 225 230 235 240 Arg Ala Pro Met Phe Gly Asp Gly Thr Thr Pro Ala Val Leu Ala Ser                 245 250 255 Ala Leu Leu Asp Ser Gly Arg His Leu Glu Ser Val Gln Ala Leu Val             260 265 270 Arg Leu Val Cys Gln Asp Pro Val His Gly Asp Gln Asn Gln Ala Thr         275 280 285 Val Arg Arg Trp Ile Glu Glu Trp Gln Pro Arg Cys Lys Ala Ala Ala     290 295 300 Gln Ser Phe Leu Pro Thr Phe Ser Asp Cys Gly Ile Asp Ala Lys Glu 305 310 315 320 Ser Ala Asn Ala Leu Ser Arg Ala Leu Ala Asn Gln Arg Ala Ala Val                 325 330 335 Glu Gly Ala Gly Ile Thr Ala             340 <210> 11 <211> 1506 <212> DNA <213> Nocardia corallina <220> <221> CDS (1) .. (1506) <400> 11 atg gca tcg aac ccc acc cag ctc cac gag aag tcg aag tcc tac gac 48 Met Ala Ser Asn Pro Thr Gln Leu His Glu Lys Ser Lys Ser Tyr Asp 1 5 10 15 tgg gac ttc acc tcc gtc gag cgg cgc ccc aag ttc gag acg aag tac 96 Trp Asp Phe Thr Ser Val Glu Arg Arg Pro Lys Phe Glu Thr Lys Tyr             20 25 30 aag atg ccc aag aag ggc aag gac ccg ttc cgc gtc ctg atc cgt gac 144 Lys Met Pro Lys Lys Gly Lys Asp Pro Phe Arg Val Leu Ile Arg Asp         35 40 45 tac atg aag atg gaa gcg gag aag gac gac cgg acc cat ggc ttc ctc 192 Tyr Met Lys Met Glu Ala Glu Lys Asp Asp Arg Thr His Gly Phe Leu     50 55 60 gac ggc gcc gtg cgg acg cgt gag gcc acc agg att gag ccg cgg ttc 240 Asp Gly Ala Val Arg Thr Arg Glu Ala Thr Arg Ile Glu Pro Arg Phe 65 70 75 80 gct gag gcc atg aag atc atg gtg ccg cag ctg acc aac gcc gag tac 288 Ala Glu Ala Met Lys Ile Met Val Pro Gln Leu Thr Asn Ala Glu Tyr                 85 90 95 cag gcg gtg gcg ggc tgc gga atg atc atc tcg gcc gtc gag aac cag 336 Gln Ala Val Ala Gly Cys Gly Met Ile Ile Ser Ala Val Glu Asn Gln             100 105 110 gag ctc cgt cag ggc tac gcc gct cag atg ctc gat gag gtg cgg cac 384 Glu Leu Arg Gln Gly Tyr Ala Ala Gln Met Leu Asp Glu Val Arg His         115 120 125 gcg cag ctc gag atg acg cta cgc aac tac tac gcg aag cac tgg tgc 432 Ala Gln Leu Glu Met Thr Leu Arg Asn Tyr Tyr Ala Lys His Trp Cys     130 135 140 gat ccc tcc ggc ttc gac atc ggt cag cgc ggc ctg tac cag cac ccc 480 Asp Pro Ser Gly Phe Asp Ile Gly Gln Arg Gly Leu Tyr Gln His Pro 145 150 155 160 gcg ggg ctg gtg tcc atc ggc gag ttc cag cac ttc aat act ggt gac 528 Ala Gly Leu Val Ser Ile Gly Glu Phe Gln His Phe Asn Thr Gly Asp                 165 170 175 ccg ctt gac gtc atc atc gat ctc aac atc gtg gcc gag acg gcg ttc 576 Pro Leu Asp Val Ile Ile Asp Leu Asn Ile Val Ala Glu Thr Ala Phe             180 185 190 acg aac atc ctg ctg gtg gcc act cca cag gtc gcc gtg gcc aac ggg 624 Thr Asn Ile Leu Leu Val Ala Thr Pro Gln Val Ala Val Ala Asn Gly         195 200 205 gac aac gcg atg gcc agc gtg ttc ctc tcg atc cag tcg gac gag gcc 672 Asp Asn Ala Met Ala Ser Val Phe Leu Ser Ile Gln Ser Asp Glu Ala     210 215 220 agg cac atg gcc aac ggg tac ggc tcg gtc atg gcg ctg ctg gag aac 720 Arg His Met Ala Asn Gly Tyr Gly Ser Val Met Ala Leu Leu Glu Asn 225 230 235 240 gag gac aac ctc ccg ctg ctc aac cag tct ctc gat cgg cac ttc tgg 768 Glu Asp Asn Leu Pro Leu Leu Asn Gln Ser Leu Asp Arg His Phe Trp                 245 250 255 cgt gcc cac aag gcc ttg gac aac gcg gtc gga tgg tgt tcg gag tat 816 Arg Ala His Lys Ala Leu Asp Asn Ala Val Gly Trp Cys Ser Glu Tyr             260 265 270 ggc gcc cgc aag cgg cca tgg agc tac aag gcc cag tgg gag gaa tgg 864 Gly Ala Arg Lys Arg Pro Trp Ser Tyr Lys Ala Gln Trp Glu Glu Trp         275 280 285 gtc gtc gac gac ttc gtg ggc ggc tac atc gac cga ctc agc gag ttc 912 Val Val Asp Asp Phe Val Gly Gly Tyr Ile Asp Arg Leu Ser Glu Phe     290 295 300 ggc gtt cag gct ccg gcc tgc ctt ggc gcg gcc gcc gac gag gtc aag 960 Gly Val Gln Ala Pro Ala Cys Leu Gly Ala Ala Ala Asp Glu Val Lys 305 310 315 320 tgg tcg cac cac acg ctc ggt cag gtg ctg tcg gcg gtg tgg ccg ctg 1008 Trp Ser His His Thr Leu Gly Gln Val Leu Ser Ala Val Trp Pro Leu                 325 330 335 aac ttc tgg cgc tcg gac gcc atg gga ccg gcg gac ttc gag tgg ttc 1056 Asn Phe Trp Arg Ser Asp Ala Met Gly Pro Ala Asp Phe Glu Trp Phe             340 345 350 gag aac cac tac ccg ggc tgg agc gcg gcc tac cag ggt tac tgg gag 1104 Glu Asn His Tyr Pro Gly Trp Ser Ala Ala Tyr Gln Gly Tyr Trp Glu         355 360 365 ggc tac aag gcg ctc gcc gac cca gca ggc gga cgc atc atg ctc cag 1152 Gly Tyr Lys Ala Leu Ala Asp Pro Ala Gly Gly Arg Ile Met Leu Gln     370 375 380 gag ctg ccg ggt ctg ccg ccg atg tgt cag gtg tgc cag gtg ccg tgc 1200 Glu Leu Pro Gly Leu Pro Pro Met Cys Gln Val Cys Gln Val Pro Cys 385 390 395 400 gtg atg ccg cgg ctg gat atg aac gcc gcg cgg atc atc gag ttc gag 1248 Val Met Pro Arg Leu Asp Met Asn Ala Ala Arg Ile Ile Glu Phe Glu                 405 410 415 ggg cag aaa atc gcg ctg tgc agc gaa ccc tgc cag cgg atc ttc acc 1296 Gly Gln Lys Ile Ala Leu Cys Ser Glu Pro Cys Gln Arg Ile Phe Thr             420 425 430 aac tgg ccg gag gcg tac cgc cac cgc aag caa tac tgg gcc cgc tac 1344 Asn Trp Pro Glu Ala Tyr Arg His Arg Lys Gln Tyr Trp Ala Arg Tyr         435 440 445 cac gga tgg gac ctg gcg gac gtc atc gtt gat ctc ggc tac atc cgc 1392 His Gly Trp Asp Leu Ala Asp Val Ile Val Asp Leu Gly Tyr Ile Arg     450 455 460 ccg gac ggc aag acc ctc atc ggc cag ccg ctg ctc gag atg gag cgg 1440 Pro Asp Gly Lys Thr Leu Ile Gly Gln Pro Leu Leu Glu Met Glu Arg 465 470 475 480 ctg tgg acc atc gac gac atc cgg gcc ctt cag tac gaa gtc aag gac 1488 Leu Trp Thr Ile Asp Asp Ile Arg Ala Leu Gln Tyr Glu Val Lys Asp                 485 490 495 ccg ttg cag gag gcg tga 1506 Pro Leu Gln Glu Ala             500 <210> 12 <211> 501 <212> PRT <213> Nocardia corallina <400> 12 Met Ala Ser Asn Pro Thr Gln Leu His Glu Lys Ser Lys Ser Tyr Asp 1 5 10 15 Trp Asp Phe Thr Ser Val Glu Arg Arg Pro Lys Phe Glu Thr Lys Tyr             20 25 30 Lys Met Pro Lys Lys Gly Lys Asp Pro Phe Arg Val Leu Ile Arg Asp         35 40 45 Tyr Met Lys Met Glu Ala Glu Lys Asp Asp Arg Thr His Gly Phe Leu     50 55 60 Asp Gly Ala Val Arg Thr Arg Glu Ala Thr Arg Ile Glu Pro Arg Phe 65 70 75 80 Ala Glu Ala Met Lys Ile Met Val Pro Gln Leu Thr Asn Ala Glu Tyr                 85 90 95 Gln Ala Val Ala Gly Cys Gly Met Ile Ile Ser Ala Val Glu Asn Gln             100 105 110 Glu Leu Arg Gln Gly Tyr Ala Ala Gln Met Leu Asp Glu Val Arg His         115 120 125 Ala Gln Leu Glu Met Thr Leu Arg Asn Tyr Tyr Ala Lys His Trp Cys     130 135 140 Asp Pro Ser Gly Phe Asp Ile Gly Gln Arg Gly Leu Tyr Gln His Pro 145 150 155 160 Ala Gly Leu Val Ser Ile Gly Glu Phe Gln His Phe Asn Thr Gly Asp                 165 170 175 Pro Leu Asp Val Ile Ile Asp Leu Asn Ile Val Ala Glu Thr Ala Phe             180 185 190 Thr Asn Ile Leu Leu Val Ala Thr Pro Gln Val Ala Val Ala Asn Gly         195 200 205 Asp Asn Ala Met Ala Ser Val Phe Leu Ser Ile Gln Ser Asp Glu Ala     210 215 220 Arg His Met Ala Asn Gly Tyr Gly Ser Val Met Ala Leu Leu Glu Asn 225 230 235 240 Glu Asp Asn Leu Pro Leu Leu Asn Gln Ser Leu Asp Arg His Phe Trp                 245 250 255 Arg Ala His Lys Ala Leu Asp Asn Ala Val Gly Trp Cys Ser Glu Tyr             260 265 270 Gly Ala Arg Lys Arg Pro Trp Ser Tyr Lys Ala Gln Trp Glu Glu Trp         275 280 285 Val Val Asp Asp Phe Val Gly Gly Tyr Ile Asp Arg Leu Ser Glu Phe     290 295 300 Gly Val Gln Ala Pro Ala Cys Leu Gly Ala Ala Ala Asp Glu Val Lys 305 310 315 320 Trp Ser His His Thr Leu Gly Gln Val Leu Ser Ala Val Trp Pro Leu                 325 330 335 Asn Phe Trp Arg Ser Asp Ala Met Gly Pro Ala Asp Phe Glu Trp Phe             340 345 350 Glu Asn His Tyr Pro Gly Trp Ser Ala Ala Tyr Gln Gly Tyr Trp Glu         355 360 365 Gly Tyr Lys Ala Leu Ala Asp Pro Ala Gly Gly Arg Ile Met Leu Gln     370 375 380 Glu Leu Pro Gly Leu Pro Pro Met Cys Gln Val Cys Gln Val Pro Cys 385 390 395 400 Val Met Pro Arg Leu Asp Met Asn Ala Ala Arg Ile Ile Glu Phe Glu                 405 410 415 Gly Gln Lys Ile Ala Leu Cys Ser Glu Pro Cys Gln Arg Ile Phe Thr             420 425 430 Asn Trp Pro Glu Ala Tyr Arg His Arg Lys Gln Tyr Trp Ala Arg Tyr         435 440 445 His Gly Trp Asp Leu Ala Asp Val Ile Val Asp Leu Gly Tyr Ile Arg     450 455 460 Pro Asp Gly Lys Thr Leu Ile Gly Gln Pro Leu Leu Glu Met Glu Arg 465 470 475 480 Leu Trp Thr Ile Asp Asp Ile Arg Ala Leu Gln Tyr Glu Val Lys Asp                 485 490 495 Pro Leu Gln Glu Ala             500 <210> 13 <211> 1494 <212> DNA <213> Xanthobacta sp. <220> <221> CDS (222) (1) .. (1494) <400> 13 atg gcg ctc ttg aat cgg gac gat tgg tac gac atc gcg cgc gat gtc 48 Met Ala Leu Leu Asn Arg Asp Asp Trp Tyr Asp Ile Ala Arg Asp Val 1 5 10 15 gac tgg acg ctc agc tat gtc gac cgc gcg gtc gcc ttt ccc gag gag 96 Asp Trp Thr Leu Ser Tyr Val Asp Arg Ala Val Ala Phe Pro Glu Glu             20 25 30 tgg aaa ggc gaa aag gac att tgc ggc acg gcc tgg gac gat tgg gac 144 Trp Lys Gly Glu Lys Asp Ile Cys Gly Thr Ala Trp Asp Asp Trp Asp         35 40 45 gag ccc ttc cgg gtc tcc ttc cgc gaa tat gtg atg gtc cag cgc gac 192 Glu Pro Phe Arg Val Ser Phe Arg Glu Tyr Val Met Val Gln Arg Asp     50 55 60 aag gaa gcg agc gtc ggc gcc atc cgc gag gcc atg gtc cgc gcc aag 240 Lys Glu Ala Ser Val Gly Ala Ile Arg Glu Ala Met Val Arg Ala Lys 65 70 75 80 gcc tat gag aag ctc gac gac ggc cac aag gcc acc tcg cac ctg cac 288 Ala Tyr Glu Lys Leu Asp Asp Gly His Lys Ala Thr Ser His Leu His                 85 90 95 atg ggc acc atc acc atg gtg gag cac atg gcg gtc acc atg cag agc 336 Met Gly Thr Ile Thr Met Val Glu His Met Ala Val Thr Met Gln Ser             100 105 110 cgg ttc gtg cgc ttc gcg ccg tcc gcc cgc tgg cgc agc ctc ggg gcg 384 Arg Phe Val Arg Phe Ala Pro Ser Ala Arg Trp Arg Ser Leu Gly Ala         115 120 125 ttc ggc atg ctg gac gag acc cgc cac acc cag ctc gac ctg cgc ttc 432 Phe Gly Met Leu Asp Glu Thr Arg His Thr Gln Leu Asp Leu Arg Phe     130 135 140 agc cac gat ctg ctc aac gat tcc ccg agc ttc gac tgg agc cag cgg 480 Ser His Asp Leu Leu Asn Asp Ser Pro Ser Phe Asp Trp Ser Gln Arg 145 150 155 160 gcg ttc cac acc gac gaa tgg gcg gtt ctc gcc acc cgc aac ctg ttc 528 Ala Phe His Thr Asp Glu Trp Ala Val Leu Ala Thr Arg Asn Leu Phe                 165 170 175 gac gac atc atg ctc aac gcc gac tgc gtg gag gcg gcg ctc gcc acc 576 Asp Asp Ile Met Leu Asn Ala Asp Cys Val Glu Ala Ala Leu Ala Thr             180 185 190 agc ctg acg ctg gag cac ggc ttc acc aac atc cag ttc gtg gcg ctc 624 Ser Leu Thr Leu Glu His Gly Phe Thr Asn Ile Gln Phe Val Ala Leu         195 200 205 gcc tcc gac gcc atg gaa gcc ggc gac gtg aac ttc tcc aac ctc ttg 672 Ala Ser Asp Ala Met Glu Ala Gly Asp Val Asn Phe Ser Asn Leu Leu     210 215 220 tcg agc atc cag acc gac gag gcg cgg cac gcc cag ttg ggc ttt ccc 720 Ser Ser Ile Gln Thr Asp Glu Ala Arg His Ala Gln Leu Gly Phe Pro 225 230 235 240 acc ctc gac gtg atg atg aag cac gac ccc aag cgc gcc cag cag atc 768 Thr Leu Asp Val Met Met Lys His Asp Pro Lys Arg Ala Gln Gln Ile                 245 250 255 ctg gac gtc gcc ttc tgg cgc tcc tac cgc atc ttc cag gcg gtg acc 816 Leu Asp Val Ala Phe Trp Arg Ser Tyr Arg Ile Phe Gln Ala Val Thr             260 265 270 ggc gtc tcc atg gac tac tac acg ccg gtc gcc aag cgg cag atg tcg 864 Gly Val Ser Met Asp Tyr Tyr Thr Pro Val Ala Lys Arg Gln Met Ser         275 280 285 ttc aag gag ttc atg ctg gag tgg atc gtc aag cat cat gag cgc atc 912 Phe Lys Glu Phe Met Leu Glu Trp Ile Val Lys His His Glu Arg Ile     290 295 300 ctg cgc gac tac ggc ctc cag aag ccc tgg tac tgg gac acg ttc gag 960 Leu Arg Asp Tyr Gly Leu Gln Lys Pro Trp Tyr Trp Asp Thr Phe Glu 305 310 315 320 aag acc ctc gat cac ggc cac cac gcg ctg cac atc ggc acc tgg ttc 1008 Lys Thr Leu Asp His Gly His His Ala Leu His Ile Gly Thr Trp Phe                 325 330 335 tgg cgc ccg acc ctg ttc tgg gat ccc aat ggc ggc gtc tcg cgc gag 1056 Trp Arg Pro Thr Leu Phe Trp Asp Pro Asn Gly Gly Val Ser Arg Glu             340 345 350 gag cgg cgc tgg ctg aac cag aag tat ccg aac tgg gaa gag agc tgg 1104 Glu Arg Arg Trp Leu Asn Gln Lys Tyr Pro Asn Trp Glu Glu Ser Trp         355 360 365 ggc gtc ctg tgg gac gag atc atc tcc aac atc aat gcg ggc aac att 1152 Gly Val Leu Trp Asp Glu Ile Ile Ser Asn Ile Asn Ala Gly Asn Ile     370 375 380 gaa aag acc ttg ccc gag acg ctg ccg atg ctg tgc aac gtc acc aac 1200 Glu Lys Thr Leu Pro Glu Thr Leu Pro Met Leu Cys Asn Val Thr Asn 385 390 395 400 ctg ccc atc ggc tcg cac tgg gac cgc ttc cac ctg aag ccc gag cag 1248 Leu Pro Ile Gly Ser His Trp Asp Arg Phe His Leu Lys Pro Glu Gln                 405 410 415 ctc gtc tac aag ggg cgg ctc tac acc ttc gac agc gac gtc tcc aag 1296 Leu Val Tyr Lys Gly Arg Leu Tyr Thr Phe Asp Ser Asp Val Ser Lys             420 425 430 tgg atc ttc gag ctc gat ccg gag cgc tat gcc ggc cac acc aac gtg 1344 Trp Ile Phe Glu Leu Asp Pro Glu Arg Tyr Ala Gly His Thr Asn Val         435 440 445 gtc gac cgc ttc atc ggc ggg cag atc cag ccc atg acc atc gag ggc 1392 Val Asp Arg Phe Ile Gly Gly Gln Ile Gln Pro Met Thr Ile Glu Gly     450 455 460 gtg ctc aac tgg atg ggc ctg acg ccc gaa gtc atg ggc aag gac gtg 1440 Val Leu Asn Trp Met Gly Leu Thr Pro Glu Val Met Gly Lys Asp Val 465 470 475 480 ttc aac tac cgt tgg gcc ggc gat tac gcc gag aac cgg atc gcc gcc 1488 Phe Asn Tyr Arg Trp Ala Gly Asp Tyr Ala Glu Asn Arg Ile Ala Ala                 485 490 495 gag taa 1494 Glu                                                                          <210> 14 <211> 497 <212> PRT <213> Xanthobacta sp. <400> 14 Met Ala Leu Leu Asn Arg Asp Asp Trp Tyr Asp Ile Ala Arg Asp Val 1 5 10 15 Asp Trp Thr Leu Ser Tyr Val Asp Arg Ala Val Ala Phe Pro Glu Glu             20 25 30 Trp Lys Gly Glu Lys Asp Ile Cys Gly Thr Ala Trp Asp Asp Trp Asp         35 40 45 Glu Pro Phe Arg Val Ser Phe Arg Glu Tyr Val Met Val Gln Arg Asp     50 55 60 Lys Glu Ala Ser Val Gly Ala Ile Arg Glu Ala Met Val Arg Ala Lys 65 70 75 80 Ala Tyr Glu Lys Leu Asp Asp Gly His Lys Ala Thr Ser His Leu His                 85 90 95 Met Gly Thr Ile Thr Met Val Glu His Met Ala Val Thr Met Gln Ser             100 105 110 Arg Phe Val Arg Phe Ala Pro Ser Ala Arg Trp Arg Ser Leu Gly Ala         115 120 125 Phe Gly Met Leu Asp Glu Thr Arg His Thr Gln Leu Asp Leu Arg Phe     130 135 140 Ser His Asp Leu Leu Asn Asp Ser Pro Ser Phe Asp Trp Ser Gln Arg 145 150 155 160 Ala Phe His Thr Asp Glu Trp Ala Val Leu Ala Thr Arg Asn Leu Phe                 165 170 175 Asp Asp Ile Met Leu Asn Ala Asp Cys Val Glu Ala Ala Leu Ala Thr             180 185 190 Ser Leu Thr Leu Glu His Gly Phe Thr Asn Ile Gln Phe Val Ala Leu         195 200 205 Ala Ser Asp Ala Met Glu Ala Gly Asp Val Asn Phe Ser Asn Leu Leu     210 215 220 Ser Ser Ile Gln Thr Asp Glu Ala Arg His Ala Gln Leu Gly Phe Pro 225 230 235 240 Thr Leu Asp Val Met Met Lys His Asp Pro Lys Arg Ala Gln Gln Ile                 245 250 255 Leu Asp Val Ala Phe Trp Arg Ser Tyr Arg Ile Phe Gln Ala Val Thr             260 265 270 Gly Val Ser Met Asp Tyr Tyr Thr Pro Val Ala Lys Arg Gln Met Ser         275 280 285 Phe Lys Glu Phe Met Leu Glu Trp Ile Val Lys His His Glu Arg Ile     290 295 300 Leu Arg Asp Tyr Gly Leu Gln Lys Pro Trp Tyr Trp Asp Thr Phe Glu 305 310 315 320 Lys Thr Leu Asp His Gly His His Ala Leu His Ile Gly Thr Trp Phe                 325 330 335 Trp Arg Pro Thr Leu Phe Trp Asp Pro Asn Gly Gly Val Ser Arg Glu             340 345 350 Glu Arg Arg Trp Leu Asn Gln Lys Tyr Pro Asn Trp Glu Glu Ser Trp         355 360 365 Gly Val Leu Trp Asp Glu Ile Ile Ser Asn Ile Asn Ala Gly Asn Ile     370 375 380 Glu Lys Thr Leu Pro Glu Thr Leu Pro Met Leu Cys Asn Val Thr Asn 385 390 395 400 Leu Pro Ile Gly Ser His Trp Asp Arg Phe His Leu Lys Pro Glu Gln                 405 410 415 Leu Val Tyr Lys Gly Arg Leu Tyr Thr Phe Asp Ser Asp Val Ser Lys             420 425 430 Trp Ile Phe Glu Leu Asp Pro Glu Arg Tyr Ala Gly His Thr Asn Val         435 440 445 Val Asp Arg Phe Ile Gly Gly Gln Ile Gln Pro Met Thr Ile Glu Gly     450 455 460 Val Leu Asn Trp Met Gly Leu Thr Pro Glu Val Met Gly Lys Asp Val 465 470 475 480 Phe Asn Tyr Arg Trp Ala Gly Asp Tyr Ala Glu Asn Arg Ile Ala Ala                 485 490 495 Glu      <210> 15 <211> 1026 <212> DNA <213> Xanthobacta sp. <220> <221> CDS (222) (1) .. (1026) <400> 15 atg aca cag cag cgc ccc acc cgc acg cgc gag cgc aag aag acc tgg 48 Met Thr Gln Gln Arg Pro Thr Arg Thr Arg Glu Arg Lys Lys Thr Trp 1 5 10 15 acg gct ttc ggc aat ctc gga cgc aag ccg acc gac tac gag gtc gtc 96 Thr Ala Phe Gly Asn Leu Gly Arg Lys Pro Thr Asp Tyr Glu Val Val             20 25 30 acc cac aac atg aac cac acc atg cgc ggc acg ccc ctg gag ctg tcg 144 Thr His Asn Met Asn His Thr Met Arg Gly Thr Pro Leu Glu Leu Ser         35 40 45 ccg acg gtg cac gcc aat gtg tgg ctc aag aag aac cgc gac gag atc 192 Pro Thr Val His Ala Asn Val Trp Leu Lys Lys Asn Arg Asp Glu Ile     50 55 60 gcg ctc aag gtc gac agc tgg gat ctg ttc cgc gat ccc gac cgc acc 240 Ala Leu Lys Val Asp Ser Trp Asp Leu Phe Arg Asp Pro Asp Arg Thr 65 70 75 80 acc tac gac acc tac gtc aag atg cag gac gac cag gag acc tat gtc 288 Thr Tyr Asp Thr Tyr Val Lys Met Gln Asp Asp Gln Glu Thr Tyr Val                 85 90 95 gac aac ctg ctc ctg tcc tac acc ggc gag ggc cgc tac gac gag gag 336 Asp Asn Leu Leu Leu Ser Tyr Thr Gly Glu Gly Arg Tyr Asp Glu Glu             100 105 110 ctt tcc tcg cgc agc ctc gac ctc ctg tcc gcg ggg ctg acg ccg acc 384 Leu Ser Ser Arg Ser Leu Asp Leu Leu Ser Ala Gly Leu Thr Pro Thr         115 120 125 cgc tat ctg ggc cat ggg ctg cag atg ctc gcg gcc tat atc cag cag 432 Arg Tyr Leu Gly His Gly Leu Gln Met Leu Ala Ala Tyr Ile Gln Gln     130 135 140 ctc gcc ccg tcg gcc tat gtg ggc aat tgc gcg gtg ttc cag acc tcc 480 Leu Ala Pro Ser Ala Tyr Val Gly Asn Cys Ala Val Phe Gln Thr Ser 145 150 155 160 gac gcg ctg cgc cgc gtg cag cgc gtc gcc tac cgc acc cgc cag ctc 528 Asp Ala Leu Arg Arg Val Gln Arg Val Ala Tyr Arg Thr Arg Gln Leu                 165 170 175 gcc gac gcc cat ccg gcc cgc ggc ttc ggc tcc ggc gac cgg gcg gtg 576 Ala Asp Ala His Pro Ala Arg Gly Phe Gly Ser Gly Asp Arg Ala Val             180 185 190 tgg gag aag tcc ccg gac tgg cag ccc atc cgc aag gcc atc gag gag 624 Trp Glu Lys Ser Pro Asp Trp Gln Pro Ile Arg Lys Ala Ile Glu Glu         195 200 205 ctg ctc gtc acc ttc gaa tgg gac aag gcg ctc gcc ggc acc aat ttc 672 Leu Leu Val Thr Phe Glu Trp Asp Lys Ala Leu Ala Gly Thr Asn Phe     210 215 220 gtg gtg aag ccg atc ctc gac gag ctg ttc ctc aac cac ctg gcg cgc 720 Val Val Lys Pro Ile Leu Asp Glu Leu Phe Leu Asn His Leu Ala Arg 225 230 235 240 ctg ctc cac gtg gag ggc gac gag ctc gac agc ctc gtg ctg cgg aac 768 Leu Leu His Val Glu Gly Asp Glu Leu Asp Ser Leu Val Leu Arg Asn                 245 250 255 ctt cac ggc gac gcc cag cgc cac gcc cgc tgg acg gcc gcg ctc ggc 816 Leu His Gly Asp Ala Gln Arg His Ala Arg Trp Thr Ala Ala Leu Gly             260 265 270 cgc ttc gcc gtc gag cag aac gtg aac aac cgc acg gtc ctg cgc gac 864 Arg Phe Ala Val Glu Gln Asn Val Asn Asn Arg Thr Val Leu Arg Asp         275 280 285 gcc atc gcc ggc tgg cac gag acc ggc gag gcg gtc ctc gcc gcg ggc 912 Ala Ile Ala Gly Trp His Glu Thr Gly Glu Ala Val Leu Ala Ala Gly     290 295 300 gcc ggg atg ctt gcg agc cgc gcc ccc agc gcg gat gcg gcc aag atc 960 Ala Gly Met Leu Ala Ser Arg Ala Pro Ser Ala Asp Ala Ala Lys Ile 305 310 315 320 gcc gac gag gtc cgc gcc acg ctc gcg cag ctg cac gcc aat gcg ggc 1008 Ala Asp Glu Val Arg Ala Thr Leu Ala Gln Leu His Ala Asn Ala Gly                 325 330 335 ctc ggg cac gat gcc tga 1026 Leu Gly His Asp Ala             340 <210> 16 <211> 341 <212> PRT <213> Xanthobacta sp. <400> 16 Met Thr Gln Gln Arg Pro Thr Arg Thr Arg Glu Arg Lys Lys Thr Trp 1 5 10 15 Thr Ala Phe Gly Asn Leu Gly Arg Lys Pro Thr Asp Tyr Glu Val Val             20 25 30 Thr His Asn Met Asn His Thr Met Arg Gly Thr Pro Leu Glu Leu Ser         35 40 45 Pro Thr Val His Ala Asn Val Trp Leu Lys Lys Asn Arg Asp Glu Ile     50 55 60 Ala Leu Lys Val Asp Ser Trp Asp Leu Phe Arg Asp Pro Asp Arg Thr 65 70 75 80 Thr Tyr Asp Thr Tyr Val Lys Met Gln Asp Asp Gln Glu Thr Tyr Val                 85 90 95 Asp Asn Leu Leu Leu Ser Tyr Thr Gly Glu Gly Arg Tyr Asp Glu Glu             100 105 110 Leu Ser Ser Arg Ser Leu Asp Leu Leu Ser Ala Gly Leu Thr Pro Thr         115 120 125 Arg Tyr Leu Gly His Gly Leu Gln Met Leu Ala Ala Tyr Ile Gln Gln     130 135 140 Leu Ala Pro Ser Ala Tyr Val Gly Asn Cys Ala Val Phe Gln Thr Ser 145 150 155 160 Asp Ala Leu Arg Arg Val Gln Arg Val Ala Tyr Arg Thr Arg Gln Leu                 165 170 175 Ala Asp Ala His Pro Ala Arg Gly Phe Gly Ser Gly Asp Arg Ala Val             180 185 190 Trp Glu Lys Ser Pro Asp Trp Gln Pro Ile Arg Lys Ala Ile Glu Glu         195 200 205 Leu Leu Val Thr Phe Glu Trp Asp Lys Ala Leu Ala Gly Thr Asn Phe     210 215 220 Val Val Lys Pro Ile Leu Asp Glu Leu Phe Leu Asn His Leu Ala Arg 225 230 235 240 Leu Leu His Val Glu Gly Asp Glu Leu Asp Ser Leu Val Leu Arg Asn                 245 250 255 Leu His Gly Asp Ala Gln Arg His Ala Arg Trp Thr Ala Ala Leu Gly             260 265 270 Arg Phe Ala Val Glu Gln Asn Val Asn Asn Arg Thr Val Leu Arg Asp         275 280 285 Ala Ile Ala Gly Trp His Glu Thr Gly Glu Ala Val Leu Ala Ala Gly     290 295 300 Ala Gly Met Leu Ala Ser Arg Ala Pro Ser Ala Asp Ala Ala Lys Ile 305 310 315 320 Ala Asp Glu Val Arg Ala Thr Leu Ala Gln Leu His Ala Asn Ala Gly                 325 330 335 Leu Gly His Asp Ala             340 <210> 17 <211> 267 <212> DNA <213> Xanthobacta sp. <220> <221> CDS (222) (1) .. (267) <400> 17 atg tct ttg ttc ccc atc gtg ggc cgc ttc gtg ggg gat ttc gtc ccc 48 Met Ser Leu Phe Pro Ile Val Gly Arg Phe Val Gly Asp Phe Val Pro 1 5 10 15 cac ctg gtg gcg gtg gac acc tct gac acc atc gat cag atc gcc gag 96 His Leu Val Ala Val Asp Thr Ser Asp Thr Ile Asp Gln Ile Ala Glu             20 25 30 aag gtg gcg gtc cac acg gtc ggg cgg cgc ttg ccg ccc gat ccc acc 144 Lys Val Ala Val His Thr Val Gly Arg Arg Leu Pro Pro Asp Pro Thr         35 40 45 gcc acc ggc tat gag gtg ctc ctc gac ggc gag acc ctg gac ggg ggc 192 Ala Thr Gly Tyr Glu Val Leu Leu Asp Gly Glu Thr Leu Asp Gly Gly     50 55 60 gcc acc ctg gag gcc atc atg acc aag cgc gag atg ctg ccc ctg cag 240 Ala Thr Leu Glu Ala Ile Met Thr Lys Arg Glu Met Leu Pro Leu Gln 65 70 75 80 tgg ttc gac gtg agg ttc aag aag tga 267 Trp Phe Asp Val Arg Phe Lys Lys                 85 <210> 18 <211> 88 <212> PRT <213> Xanthobacta sp. <400> 18 Met Ser Leu Phe Pro Ile Val Gly Arg Phe Val Gly Asp Phe Val Pro 1 5 10 15 His Leu Val Ala Val Asp Thr Ser Asp Thr Ile Asp Gln Ile Ala Glu             20 25 30 Lys Val Ala Val His Thr Val Gly Arg Arg Leu Pro Pro Asp Pro Thr         35 40 45 Ala Thr Gly Tyr Glu Val Leu Leu Asp Gly Glu Thr Leu Asp Gly Gly     50 55 60 Ala Thr Leu Glu Ala Ile Met Thr Lys Arg Glu Met Leu Pro Leu Gln 65 70 75 80 Trp Phe Asp Val Arg Phe Lys Lys                 85 <210> 19 <211> 1584 <212> DNA <213> Methylococcus capsulatas <220> <221> CDS (222) (1) .. (1584) <400> 19 atg gca ctt agc acc gca acc aag gcc gcg acg gac gcg ctg gct gcc 48 Met Ala Leu Ser Thr Ala Thr Lys Ala Ala Thr Asp Ala Leu Ala Ala 1 5 10 15 aat cgg gca ccc acc agc gtg aat gca cag gaa gtg cac cgt tgg ctc 96 Asn Arg Ala Pro Thr Ser Val Asn Ala Gln Glu Val His Arg Trp Leu             20 25 30 cag agc ttc aac tgg gat ttc aag aac aac cgg acc aag tac gcc acc 144 Gln Ser Phe Asn Trp Asp Phe Lys Asn Asn Arg Thr Lys Tyr Ala Thr         35 40 45 aag tac aag atg gcg aac gag acc aag gaa cag ttc aag ctg atc gcc 192 Lys Tyr Lys Met Ala Asn Glu Thr Lys Glu Gln Phe Lys Leu Ile Ala     50 55 60 aag gaa tat gcg cgc atg gag gca gtc aag gac gaa agg cag ttc ggt 240 Lys Glu Tyr Ala Arg Met Glu Ala Val Lys Asp Glu Arg Gln Phe Gly 65 70 75 80 agc ctg cag gat gcg ctg acc cgc ctc aac gcc ggt gtt cgc gtt cat 288 Ser Leu Gln Asp Ala Leu Thr Arg Leu Asn Ala Gly Val Arg Val His                 85 90 95 ccg aag tgg aac gag acc atg aaa gtg gtt tcg aac ttc ctg gaa gtg 336 Pro Lys Trp Asn Glu Thr Met Lys Val Val Ser Asn Phe Leu Glu Val             100 105 110 ggc gaa tac aac gcc atc gcc gct acc ggg atg ctg tgg gat tcc gcc 384 Gly Glu Tyr Asn Ala Ile Ala Ala Thr Gly Met Leu Trp Asp Ser Ala         115 120 125 cag gcg gcg gaa cag aag aac ggc tat ctg gcc cag gtg ttg gat gaa 432 Gln Ala Ala Glu Gln Lys Asn Gly Tyr Leu Ala Gln Val Leu Asp Glu     130 135 140 atc cgc cac acc cac cag tgt gcc tac gtc aac tac tac ttc gcg aag 480 Ile Arg His Thr His Gln Cys Ala Tyr Val Asn Tyr Tyr Phe Ala Lys 145 150 155 160 aac ggc cag gac ccg gcc ggt cac aac gat gct cgc cgc acc cgt acc 528 Asn Gly Gln Asp Pro Ala Gly His Asn Asp Ala Arg Arg Thr Arg Thr                 165 170 175 atc ggt ccg ctg tgg aag ggc atg aag cgc gtg ttt tcc gac ggc ttc 576 Ile Gly Pro Leu Trp Lys Gly Met Lys Arg Val Phe Ser Asp Gly Phe             180 185 190 att tcc ggc gac gcc gtg gaa tgc tcc ctc aac ctg cag ctg gtg ggt 624 Ile Ser Gly Asp Ala Val Glu Cys Ser Leu Asn Leu Gln Leu Val Gly         195 200 205 gag gcc tgc ttc acc aat ccg ctg atc gtc gca gtg acc gaa tgg gct 672 Glu Ala Cys Phe Thr Asn Pro Leu Ile Val Ala Val Thr Glu Trp Ala     210 215 220 gcc gcc aac ggc gat gaa atc acc ccg acg gtg ttc ctg tcg atc gag 720 Ala Ala Asn Gly Asp Glu Ile Thr Pro Thr Val Phe Leu Ser Ile Glu 225 230 235 240 acc gac gaa ctg cgc cac atg gcc aac ggt tac cag acc gtc gtt tcc 768 Thr Asp Glu Leu Arg His Met Ala Asn Gly Tyr Gln Thr Val Val Ser                 245 250 255 atc gcc aac gat ccg gct tcc gcc aag tat ctc aac acg gac ctg aac 816 Ile Ala Asn Asp Pro Ala Ser Ala Lys Tyr Leu Asn Thr Asp Leu Asn             260 265 270 aac gcc ttc tgg acc cag cag aag tac ttc acg ccg gtg ttg ggc atg 864 Asn Ala Phe Trp Thr Gln Gln Lys Tyr Phe Thr Pro Val Leu Gly Met         275 280 285 ctg ttc gag tat ggc tcc aag ttc aag gtc gag ccg tgg gtc aag acg 912 Leu Phe Glu Tyr Gly Ser Lys Phe Lys Val Glu Pro Trp Val Lys Thr     290 295 300 tgg gac cgc tgg gtg tac gag gac tgg ggc ggc atc tgg atc ggc cgt 960 Trp Asp Arg Trp Val Tyr Glu Asp Trp Gly Gly Ile Trp Ile Gly Arg 305 310 315 320 ctg ggc aag tac ggg gtg gag tcg ccg cgc agc ctc aag gac gcc aag 1008 Leu Gly Lys Tyr Gly Val Glu Ser Pro Arg Ser Leu Lys Asp Ala Lys                 325 330 335 cag gac gct tac tgg gct cac cac gac ctg tat ctg ctg gct tat gcg 1056 Gln Asp Ala Tyr Trp Ala His His Asp Leu Tyr Leu Leu Ala Tyr Ala             340 345 350 ctg tgg ccg acc ggc ttc ttc cgt ctg gcg ctg ccg gat cag gaa gaa 1104 Leu Trp Pro Thr Gly Phe Phe Arg Leu Ala Leu Pro Asp Gln Glu Glu         355 360 365 atg gag tgg ttc gag gcc aac tac ccc ggc tgg tac gac cac tac ggc 1152 Met Glu Trp Phe Glu Ala Asn Tyr Pro Gly Trp Tyr Asp His Tyr Gly     370 375 380 aag atc tac gag gaa tgg cgc gcc cgc ggt tgc gag gat ccg tcc tcg 1200 Lys Ile Tyr Glu Glu Trp Arg Ala Arg Gly Cys Glu Asp Pro Ser Ser 385 390 395 400 ggc ttc atc ccg ctg atg tgg ttc atc gaa aac aac cat ccc atc tac 1248 Gly Phe Ile Pro Leu Met Trp Phe Ile Glu Asn Asn His Pro Ile Tyr                 405 410 415 atc gat cgc gtg tcg caa gtg ccg ttc tgc ccg agc ttg gcc aag ggc 1296 Ile Asp Arg Val Ser Gln Val Pro Phe Cys Pro Ser Leu Ala Lys Gly             420 425 430 gcc agc acc ctg cgc gtg cac gag tac aac ggc gag atg cac acc ttc 1344 Ala Ser Thr Leu Arg Val His Glu Tyr Asn Gly Glu Met His Thr Phe         435 440 445 agc gac cag tgg ggc gag cgc atg tgg ctg gcc gag ccg gag cgc tac 1392 Ser Asp Gln Trp Gly Glu Arg Met Trp Leu Ala Glu Pro Glu Arg Tyr     450 455 460 gag tgc cag aac atc ttc gaa cag tac gaa gga cgc gaa ctg tcg gaa 1440 Glu Cys Gln Asn Ile Phe Glu Gln Tyr Glu Gly Arg Glu Leu Ser Glu 465 470 475 480 gtg atc gcc gaa ctg cac ggg ctg cgc agt gat ggc aag acc ctg atc 1488 Val Ile Ala Glu Leu His Gly Leu Arg Ser Asp Gly Lys Thr Leu Ile                 485 490 495 gcc cag ccg cat gtc cgt ggc gac aag ctg tgg acg ttg gac gat atc 1536 Ala Gln Pro His Val Arg Gly Asp Lys Leu Trp Thr Leu Asp Asp Ile             500 505 510 aaa cgc ctg aac tgc gtc ttc aag aac ccg gtg aag gca ttc aat tga 1584 Lys Arg Leu Asn Cys Val Phe Lys Asn Pro Val Lys Ala Phe Asn         515 520 525 <210> 20 <211> 527 <212> PRT <213> Methylococcus capsulatas <400> 20 Met Ala Leu Ser Thr Ala Thr Lys Ala Ala Thr Asp Ala Leu Ala Ala 1 5 10 15 Asn Arg Ala Pro Thr Ser Val Asn Ala Gln Glu Val His Arg Trp Leu             20 25 30 Gln Ser Phe Asn Trp Asp Phe Lys Asn Asn Arg Thr Lys Tyr Ala Thr         35 40 45 Lys Tyr Lys Met Ala Asn Glu Thr Lys Glu Gln Phe Lys Leu Ile Ala     50 55 60 Lys Glu Tyr Ala Arg Met Glu Ala Val Lys Asp Glu Arg Gln Phe Gly 65 70 75 80 Ser Leu Gln Asp Ala Leu Thr Arg Leu Asn Ala Gly Val Arg Val His                 85 90 95 Pro Lys Trp Asn Glu Thr Met Lys Val Val Ser Asn Phe Leu Glu Val             100 105 110 Gly Glu Tyr Asn Ala Ile Ala Ala Thr Gly Met Leu Trp Asp Ser Ala         115 120 125 Gln Ala Ala Glu Gln Lys Asn Gly Tyr Leu Ala Gln Val Leu Asp Glu     130 135 140 Ile Arg His Thr His Gln Cys Ala Tyr Val Asn Tyr Tyr Phe Ala Lys 145 150 155 160 Asn Gly Gln Asp Pro Ala Gly His Asn Asp Ala Arg Arg Thr Arg Thr                 165 170 175 Ile Gly Pro Leu Trp Lys Gly Met Lys Arg Val Phe Ser Asp Gly Phe             180 185 190 Ile Ser Gly Asp Ala Val Glu Cys Ser Leu Asn Leu Gln Leu Val Gly         195 200 205 Glu Ala Cys Phe Thr Asn Pro Leu Ile Val Ala Val Thr Glu Trp Ala     210 215 220 Ala Ala Asn Gly Asp Glu Ile Thr Pro Thr Val Phe Leu Ser Ile Glu 225 230 235 240 Thr Asp Glu Leu Arg His Met Ala Asn Gly Tyr Gln Thr Val Val Ser                 245 250 255 Ile Ala Asn Asp Pro Ala Ser Ala Lys Tyr Leu Asn Thr Asp Leu Asn             260 265 270 Asn Ala Phe Trp Thr Gln Gln Lys Tyr Phe Thr Pro Val Leu Gly Met         275 280 285 Leu Phe Glu Tyr Gly Ser Lys Phe Lys Val Glu Pro Trp Val Lys Thr     290 295 300 Trp Asp Arg Trp Val Tyr Glu Asp Trp Gly Gly Ile Trp Ile Gly Arg 305 310 315 320 Leu Gly Lys Tyr Gly Val Glu Ser Pro Arg Ser Leu Lys Asp Ala Lys                 325 330 335 Gln Asp Ala Tyr Trp Ala His His Asp Leu Tyr Leu Leu Ala Tyr Ala             340 345 350 Leu Trp Pro Thr Gly Phe Phe Arg Leu Ala Leu Pro Asp Gln Glu Glu         355 360 365 Met Glu Trp Phe Glu Ala Asn Tyr Pro Gly Trp Tyr Asp His Tyr Gly     370 375 380 Lys Ile Tyr Glu Glu Trp Arg Ala Arg Gly Cys Glu Asp Pro Ser Ser 385 390 395 400 Gly Phe Ile Pro Leu Met Trp Phe Ile Glu Asn Asn His Pro Ile Tyr                 405 410 415 Ile Asp Arg Val Ser Gln Val Pro Phe Cys Pro Ser Leu Ala Lys Gly             420 425 430 Ala Ser Thr Leu Arg Val His Glu Tyr Asn Gly Glu Met His Thr Phe         435 440 445 Ser Asp Gln Trp Gly Glu Arg Met Trp Leu Ala Glu Pro Glu Arg Tyr     450 455 460 Glu Cys Gln Asn Ile Phe Glu Gln Tyr Glu Gly Arg Glu Leu Ser Glu 465 470 475 480 Val Ile Ala Glu Leu His Gly Leu Arg Ser Asp Gly Lys Thr Leu Ile                 485 490 495 Ala Gln Pro His Val Arg Gly Asp Lys Leu Trp Thr Leu Asp Asp Ile             500 505 510 Lys Arg Leu Asn Cys Val Phe Lys Asn Pro Val Lys Ala Phe Asn         515 520 525 <210> 21 <211> 1170 <212> DNA <213> Methylococcus capsulatas <220> <221> CDS (222) (1) .. (1170) <400> 21 atg agc atg tta gga gaa aga cgc cgc ggt ctg acc gat ccg gaa atg 48 Met Ser Met Leu Gly Glu Arg Arg Arg Gly Leu Thr Asp Pro Glu Met 1 5 10 15 gcg gcc gtc att ttg aag gcg ctt cct gaa gct ccg ctg gac ggc aac 96 Ala Ala Val Ile Leu Lys Ala Leu Pro Glu Ala Pro Leu Asp Gly Asn             20 25 30 aac aag atg ggt tat ttc gtc acc ccc cgc tgg aaa cgc ttg acg gaa 144 Asn Lys Met Gly Tyr Phe Val Thr Pro Arg Trp Lys Arg Leu Thr Glu         35 40 45 tat gaa gcc ctg acc gtt tat gcg cag ccc aac gcc gac tgg atc gcc 192 Tyr Glu Ala Leu Thr Val Tyr Ala Gln Pro Asn Ala Asp Trp Ile Ala     50 55 60 ggc ggc ctg gac tgg ggc gac tgg acc cag aaa ttc cac ggc ggc cgc 240 Gly Gly Leu Asp Trp Gly Asp Trp Thr Gln Lys Phe His Gly Gly Arg 65 70 75 80 cct tcc tgg ggc aac gag acc acg gag ctg cgc acc gtc gac tgg ttc 288 Pro Ser Trp Gly Asn Glu Thr Thr Glu Leu Arg Thr Val Asp Trp Phe                 85 90 95 aag cac cgt gac ccg ctc cgc cgt tgg cat gcg ccg tac gtc aag gac 336 Lys His Arg Asp Pro Leu Arg Arg Trp His Ala Pro Tyr Val Lys Asp             100 105 110 aag gcc gag gaa tgg cgc tac acc gac cgc ttc ctg cag ggt tac tcc 384 Lys Ala Glu Glu Trp Arg Tyr Thr Asp Arg Phe Leu Gln Gly Tyr Ser         115 120 125 gcc gac ggt cag atc cgg gcg atg aac ccg acc tgg cgg gac gag ttc 432 Ala Asp Gly Gln Ile Arg Ala Met Asn Pro Thr Trp Arg Asp Glu Phe     130 135 140 atc aac cgg tat tgg ggc gcc ttc ctg ttc aac gaa tac gga ttg ttc 480 Ile Asn Arg Tyr Trp Gly Ala Phe Leu Phe Asn Glu Tyr Gly Leu Phe 145 150 155 160 aac gct cat tcg cag ggc gcc cgg gag gcg ctg tcg gac gta acc cgc 528 Asn Ala His Ser Gln Gly Ala Arg Glu Ala Leu Ser Asp Val Thr Arg                 165 170 175 gtc agc ctg gct ttc tgg ggc ttc gac aag atc gac atc gcc cag atg 576 Val Ser Leu Ala Phe Trp Gly Phe Asp Lys Ile Asp Ile Ala Gln Met             180 185 190 atc caa ctc gaa cgg ggt ttc ctc gcc aag atc gta ccc ggt ttc gac 624 Ile Gln Leu Glu Arg Gly Phe Leu Ala Lys Ile Val Pro Gly Phe Asp         195 200 205 gag tcc aca gcg gtg ccg aag gcc gaa tgg acg aac ggg gag gtc tac 672 Glu Ser Thr Ala Val Pro Lys Ala Glu Trp Thr Asn Gly Glu Val Tyr     210 215 220 aag agc gcc cgt ctg gcc gtg gaa ggg ctg tgg cag gag gtg ttc gac 720 Lys Ser Ala Arg Leu Ala Val Glu Gly Leu Trp Gln Glu Val Phe Asp 225 230 235 240 tgg aac gag agc gct ttc tcg gtg cac gcc gtc tat gac gcg ctg ttc 768 Trp Asn Glu Ser Ala Phe Ser Val His Ala Val Tyr Asp Ala Leu Phe                 245 250 255 ggt cag ttc gtc cgc cgc gag ttc ttt cag cgg ctg gct ccc cgc ttc 816 Gly Gln Phe Val Arg Arg Glu Phe Phe Gln Arg Leu Ala Pro Arg Phe             260 265 270 ggc gac aat ctg acg cca ttc ttc atc aac cag gcc cag aca tac ttc 864 Gly Asp Asn Leu Thr Pro Phe Ile Asn Gln Ala Gln Thr Tyr Phe         275 280 285 cag atc gcc aag cag ggc gta cag gat ctg tat tac aac tgt ctg ggt 912 Gln Ile Ala Lys Gln Gly Val Gln Asp Leu Tyr Tyr Asn Cys Leu Gly     290 295 300 gac gat ccg gag ttc agc gat tac aac cgt acc gtg atg cgc aac tgg 960 Asp Asp Pro Glu Phe Ser Asp Tyr Asn Arg Thr Val Met Arg Asn Trp 305 310 315 320 acc ggc aag tgg ctg gag ccc acg atc gcc gct ctg cgc gac ttc atg 1008 Thr Gly Lys Trp Leu Glu Pro Thr Ile Ala Ala Leu Arg Asp Phe Met                 325 330 335 ggg ctg ttt gcg aag ctg ccg gcg ggc acc act gac aag gaa gaa atc 1056 Gly Leu Phe Ala Lys Leu Pro Ala Gly Thr Thr Asp Lys Glu Glu Ile             340 345 350 acc gcg tcc ctg tac cgg gtg gtc gac gac tgg atc gag gac tac gcc 1104 Thr Ala Ser Leu Tyr Arg Val Val Asp Asp Trp Ile Glu Asp Tyr Ala         355 360 365 agc gcg atc gac ttc aag gcg gac cgc gat cag atc gtt aaa gcg gtt 1152 Ser Ala Ile Asp Phe Lys Ala Asp Arg Asp Gln Ile Val Lys Ala Val     370 375 380 ctg gca gga ttg aaa taa 1170 Leu Ala Gly Leu Lys 385 <210> 22 <211> 389 <212> PRT <213> Methylococcus capsulatas <400> 22 Met Ser Met Leu Gly Glu Arg Arg Arg Gly Leu Thr Asp Pro Glu Met 1 5 10 15 Ala Ala Val Ile Leu Lys Ala Leu Pro Glu Ala Pro Leu Asp Gly Asn             20 25 30 Asn Lys Met Gly Tyr Phe Val Thr Pro Arg Trp Lys Arg Leu Thr Glu         35 40 45 Tyr Glu Ala Leu Thr Val Tyr Ala Gln Pro Asn Ala Asp Trp Ile Ala     50 55 60 Gly Gly Leu Asp Trp Gly Asp Trp Thr Gln Lys Phe His Gly Gly Arg 65 70 75 80 Pro Ser Trp Gly Asn Glu Thr Thr Glu Leu Arg Thr Val Asp Trp Phe                 85 90 95 Lys His Arg Asp Pro Leu Arg Arg Trp His Ala Pro Tyr Val Lys Asp             100 105 110 Lys Ala Glu Glu Trp Arg Tyr Thr Asp Arg Phe Leu Gln Gly Tyr Ser         115 120 125 Ala Asp Gly Gln Ile Arg Ala Met Asn Pro Thr Trp Arg Asp Glu Phe     130 135 140 Ile Asn Arg Tyr Trp Gly Ala Phe Leu Phe Asn Glu Tyr Gly Leu Phe 145 150 155 160 Asn Ala His Ser Gln Gly Ala Arg Glu Ala Leu Ser Asp Val Thr Arg                 165 170 175 Val Ser Leu Ala Phe Trp Gly Phe Asp Lys Ile Asp Ile Ala Gln Met             180 185 190 Ile Gln Leu Glu Arg Gly Phe Leu Ala Lys Ile Val Pro Gly Phe Asp         195 200 205 Glu Ser Thr Ala Val Pro Lys Ala Glu Trp Thr Asn Gly Glu Val Tyr     210 215 220 Lys Ser Ala Arg Leu Ala Val Glu Gly Leu Trp Gln Glu Val Phe Asp 225 230 235 240 Trp Asn Glu Ser Ala Phe Ser Val His Ala Val Tyr Asp Ala Leu Phe                 245 250 255 Gly Gln Phe Val Arg Arg Glu Phe Phe Gln Arg Leu Ala Pro Arg Phe             260 265 270 Gly Asp Asn Leu Thr Pro Phe Ile Asn Gln Ala Gln Thr Tyr Phe         275 280 285 Gln Ile Ala Lys Gln Gly Val Gln Asp Leu Tyr Tyr Asn Cys Leu Gly     290 295 300 Asp Asp Pro Glu Phe Ser Asp Tyr Asn Arg Thr Val Met Arg Asn Trp 305 310 315 320 Thr Gly Lys Trp Leu Glu Pro Thr Ile Ala Ala Leu Arg Asp Phe Met                 325 330 335 Gly Leu Phe Ala Lys Leu Pro Ala Gly Thr Thr Asp Lys Glu Glu Ile             340 345 350 Thr Ala Ser Leu Tyr Arg Val Val Asp Asp Trp Ile Glu Asp Tyr Ala         355 360 365 Ser Ala Ile Asp Phe Lys Ala Asp Arg Asp Gln Ile Val Lys Ala Val     370 375 380 Leu Ala Gly Leu Lys 385 <210> 23 <211> 513 <212> DNA <213> Methylococcus capsulatas <220> <221> CDS (222) (1) .. (513) <400> 23 atg gcg aaa ctg ggt ata cac agc aac gac acc cgc gac gcc tgg gtg 48 Met Ala Lys Leu Gly Ile His Ser Asn Asp Thr Arg Asp Ala Trp Val 1 5 10 15 aac aag atc gcg cag ctc aac acc ctg gaa aaa gcg gcc gag atg ctg 96 Asn Lys Ile Ala Gln Leu Asn Thr Leu Glu Lys Ala Ala Glu Met Leu             20 25 30 aag cag ttc cgg atg gac cac acc acg ccg ttc cgc aac agc tac gaa 144 Lys Gln Phe Arg Met Asp His Thr Thr Pro Phe Arg Asn Ser Tyr Glu         35 40 45 ctg gac aac gac tac ctc tgg atc gag gcc aag ctc gaa gag aag gtc 192 Leu Asp Asn Asp Tyr Leu Trp Ile Glu Ala Lys Leu Glu Glu Lys Val     50 55 60 gcc gtc ctc aag gca cgc gcc ttc aac gag gtg gac ttc cgt cat aag 240 Ala Val Leu Lys Ala Arg Ala Phe Asn Glu Val Asp Phe Arg His Lys 65 70 75 80 acc gct ttc ggc gag gat gcc aag tcc gtt ctg gac ggc acc gtc gcg 288 Thr Ala Phe Gly Glu Asp Ala Lys Ser Val Leu Asp Gly Thr Val Ala                 85 90 95 aag atg aac gcg gcc aag gac aag tgg gag gcg gag aag atc cat atc 336 Lys Met Asn Ala Ala Lys Asp Lys Trp Glu Ala Glu Lys Ile His Ile             100 105 110 ggt ttc cgc cag gcc tac aag ccg ccg atc atg ccg gtg aac tat ttc 384 Gly Phe Arg Gln Ala Tyr Lys Pro Pro Ile Met Pro Val Asn Tyr Phe         115 120 125 ctg gac ggc gag cgt cag ttg ggg acc cgg ctg atg gaa ctg cgc aac 432 Leu Asp Gly Glu Arg Gln Leu Gly Thr Arg Leu Met Glu Leu Arg Asn     130 135 140 ctc aac tac tac gac acg ccg ctg gaa gaa ctg cgc aaa cag cgc ggt 480 Leu Asn Tyr Tyr Asp Thr Pro Leu Glu Glu Leu Arg Lys Gln Arg Gly 145 150 155 160 gtg cgg gtg gtg cat ctg cag tcg ccg cac tga 513 Val Arg Val Val His Leu Gln Ser Pro His                 165 170 <210> 24 <211> 170 <212> PRT <213> Methylococcus capsulatas <400> 24 Met Ala Lys Leu Gly Ile His Ser Asn Asp Thr Arg Asp Ala Trp Val 1 5 10 15 Asn Lys Ile Ala Gln Leu Asn Thr Leu Glu Lys Ala Ala Glu Met Leu             20 25 30 Lys Gln Phe Arg Met Asp His Thr Thr Pro Phe Arg Asn Ser Tyr Glu         35 40 45 Leu Asp Asn Asp Tyr Leu Trp Ile Glu Ala Lys Leu Glu Glu Lys Val     50 55 60 Ala Val Leu Lys Ala Arg Ala Phe Asn Glu Val Asp Phe Arg His Lys 65 70 75 80 Thr Ala Phe Gly Glu Asp Ala Lys Ser Val Leu Asp Gly Thr Val Ala                 85 90 95 Lys Met Asn Ala Ala Lys Asp Lys Trp Glu Ala Glu Lys Ile His Ile             100 105 110 Gly Phe Arg Gln Ala Tyr Lys Pro Pro Ile Met Pro Val Asn Tyr Phe         115 120 125 Leu Asp Gly Glu Arg Gln Leu Gly Thr Arg Leu Met Glu Leu Arg Asn     130 135 140 Leu Asn Tyr Tyr Asp Thr Pro Leu Glu Glu Leu Arg Lys Gln Arg Gly 145 150 155 160 Val Arg Val Val His Leu Gln Ser Pro His                 165 170 <210> 25 <211> 1206 <212> DNA <213> Pseudomonas oleovorans <220> <221> CDS (222) (1) .. (1206) <400> 25 atg ctt gag aaa cac aga gtt ctg gat tcc gct cca gag tac gta gat 48 Met Leu Glu Lys His Arg Val Leu Asp Ser Ala Pro Glu Tyr Val Asp 1 5 10 15 aaa aag aaa tat ctc tgg ata cta tca act ttg tgg ccg gct act ccg 96 Lys Lys Lys Tyr Leu Trp Ile Leu Ser Thr Leu Trp Pro Ala Thr Pro             20 25 30 atg atc gga atc tgg ctt gca aat gaa act ggt tgg ggg att ttt tat 144 Met Ile Gly Ile Trp Leu Ala Asn Glu Thr Gly Trp Gly Ile Phe Tyr         35 40 45 ggg ctg gta ttg ctc gta tgg tac ggc gca ctt cca ttg ctt gat gcg 192 Gly Leu Val Leu Leu Val Trp Tyr Gly Ala Leu Pro Leu Leu Asp Ala     50 55 60 atg ttt ggt gag gac ttt aat aat ccg cct gaa gaa gtg gtg ccg aaa 240 Met Phe Gly Glu Asp Phe Asn Asn Pro Pro Glu Glu Val Val Pro Lys 65 70 75 80 cta gag aag gag cgg tac tat cga gtt ttg aca tat cta aca gtt cct 288 Leu Glu Lys Glu Arg Tyr Tyr Arg Val Leu Thr Tyr Leu Thr Val Pro                 85 90 95 atg cat tac gct gca tta att gtg tca gca tgg tgg gtc gga act cag 336 Met His Tyr Ala Ala Leu Ile Val Ser Ala Trp Trp Val Gly Thr Gln             100 105 110 cca atg tct tgg ctt gaa att ggt gcg ctt gcc ttg tca ctg ggt atc 384 Pro Met Ser Trp Leu Glu Ile Gly Ala Leu Ala Leu Ser Leu Gly Ile         115 120 125 gtg aac gga cta gcg ctc aat aca gga cac gaa ctc ggt cac aag aag 432 Val Asn Gly Leu Ala Leu Asn Thr Gly His Glu Leu Gly His Lys Lys     130 135 140 gag act ttt gat cgt tgg atg gcc aaa att gtg ttg gct gtc gta ggg 480 Glu Thr Phe Asp Arg Trp Met Ala Lys Ile Val Leu Ala Val Val Gly 145 150 155 160 tac ggt cac ttc ttt att gag cat aat aag ggt cat cac cgt gat gtc 528 Tyr Gly His Phe Phe Ile Glu His Asn Lys Gly His His Arg Asp Val                 165 170 175 gct aca ccg atg gat cct gca aca tcc cgg atg gga gaa agc att tat 576 Ala Thr Pro Met Asp Pro Ala Thr Ser Arg Met Gly Glu Ser Ile Tyr             180 185 190 aag ttt tca atc cgt gag atc cca gga gca ttt att cgt gct tgg ggg 624 Lys Phe Ser Ile Arg Glu Ile Pro Gly Ala Phe Ile Arg Ala Trp Gly         195 200 205 ctt gag gaa caa cgc ctt tcg cgc cgt ggc caa agc gtt tgg agt ttc 672 Leu Glu Glu Gln Arg Leu Ser Arg Arg Gly Gln Ser Val Trp Ser Phe     210 215 220 gat aat gaa atc ctc caa cca atg atc atc aca gtt att ctt tac gcc 720 Asp Asn Glu Ile Leu Gln Pro Met Ile Ile Thr Val Ile Leu Tyr Ala 225 230 235 240 gtt ctc ctt gcc ttg ttt gga cct aag atg ctg gtg ttc ctg ccg att 768 Val Leu Leu Ala Leu Phe Gly Pro Lys Met Leu Val Phe Leu Pro Ile                 245 250 255 caa atg gct ttc ggt tgg tgg cag ctg acc agt gcg aac tat att gaa 816 Gln Met Ala Phe Gly Trp Trp Gln Leu Thr Ser Ala Asn Tyr Ile Glu             260 265 270 cat tac ggc ttg ctc cgt caa aaa atg gag gac ggt cga tat gag cat 864 His Tyr Gly Leu Leu Arg Gln Lys Met Glu Asp Gly Arg Tyr Glu His         275 280 285 caa aag ccg cac cat tct tgg aat agt aat cac atc gtc tct aat cta 912 Gln Lys Pro His His Ser Trp Asn Ser Asn His Ile Val Ser Asn Leu     290 295 300 gtg ctg ttc cac ctt cag cgg cac tcg gat cac cac gcg cat cca aca 960 Val Leu Phe His Leu Gln Arg His Ser Asp His His Ala His Pro Thr 305 310 315 320 cgt tct tat cag tca ctt cgg gat ttt ccc ggc ctg ccg gct ctt ccg 1008 Arg Ser Tyr Gln Ser Leu Arg Asp Phe Pro Gly Leu Pro Ala Leu Pro                 325 330 335 acg ggt tac cct ggt gca ttt ttg atg gcg atg att cct cag tgg ttt 1056 Thr Gly Tyr Pro Gly Ala Phe Leu Met Ala Met Ile Pro Gln Trp Phe             340 345 350 aga tca gtt atg gat ccc aag gta gta gat tgg gct ggt ggt gac ctt 1104 Arg Ser Val Met Asp Pro Lys Val Val Asp Trp Ala Gly Gly Asp Leu         355 360 365 aat aag atc caa att gat gat tcg atg cga gaa acc tat ttg aaa aaa 1152 Asn Lys Ile Gln Ile Asp Asp Ser Met Arg Glu Thr Tyr Leu Lys Lys     370 375 380 ttt ggc act agt agt gct ggt cat agt tcg agt acc tct gcg gta gca 1200 Phe Gly Thr Ser Ser Ala Gly His Ser Ser Ser Thr Ser Ala Val Ala 385 390 395 400 tcg tag 1206 Ser                                                                          <210> 26 <211> 401 <212> PRT <213> Pseudomonas oleovorans <400> 26 Met Leu Glu Lys His Arg Val Leu Asp Ser Ala Pro Glu Tyr Val Asp 1 5 10 15 Lys Lys Lys Tyr Leu Trp Ile Leu Ser Thr Leu Trp Pro Ala Thr Pro             20 25 30 Met Ile Gly Ile Trp Leu Ala Asn Glu Thr Gly Trp Gly Ile Phe Tyr         35 40 45 Gly Leu Val Leu Leu Val Trp Tyr Gly Ala Leu Pro Leu Leu Asp Ala     50 55 60 Met Phe Gly Glu Asp Phe Asn Asn Pro Pro Glu Glu Val Val Pro Lys 65 70 75 80 Leu Glu Lys Glu Arg Tyr Tyr Arg Val Leu Thr Tyr Leu Thr Val Pro                 85 90 95 Met His Tyr Ala Ala Leu Ile Val Ser Ala Trp Trp Val Gly Thr Gln             100 105 110 Pro Met Ser Trp Leu Glu Ile Gly Ala Leu Ala Leu Ser Leu Gly Ile         115 120 125 Val Asn Gly Leu Ala Leu Asn Thr Gly His Glu Leu Gly His Lys Lys     130 135 140 Glu Thr Phe Asp Arg Trp Met Ala Lys Ile Val Leu Ala Val Val Gly 145 150 155 160 Tyr Gly His Phe Phe Ile Glu His Asn Lys Gly His His Arg Asp Val                 165 170 175 Ala Thr Pro Met Asp Pro Ala Thr Ser Arg Met Gly Glu Ser Ile Tyr             180 185 190 Lys Phe Ser Ile Arg Glu Ile Pro Gly Ala Phe Ile Arg Ala Trp Gly         195 200 205 Leu Glu Glu Gln Arg Leu Ser Arg Arg Gly Gln Ser Val Trp Ser Phe     210 215 220 Asp Asn Glu Ile Leu Gln Pro Met Ile Ile Thr Val Ile Leu Tyr Ala 225 230 235 240 Val Leu Leu Ala Leu Phe Gly Pro Lys Met Leu Val Phe Leu Pro Ile                 245 250 255 Gln Met Ala Phe Gly Trp Trp Gln Leu Thr Ser Ala Asn Tyr Ile Glu             260 265 270 His Tyr Gly Leu Leu Arg Gln Lys Met Glu Asp Gly Arg Tyr Glu His         275 280 285 Gln Lys Pro His His Ser Trp Asn Ser Asn His Ile Val Ser Asn Leu     290 295 300 Val Leu Phe His Leu Gln Arg His Ser Asp His His Ala His Pro Thr 305 310 315 320 Arg Ser Tyr Gln Ser Leu Arg Asp Phe Pro Gly Leu Pro Ala Leu Pro                 325 330 335 Thr Gly Tyr Pro Gly Ala Phe Leu Met Ala Met Ile Pro Gln Trp Phe             340 345 350 Arg Ser Val Met Asp Pro Lys Val Val Asp Trp Ala Gly Gly Asp Leu         355 360 365 Asn Lys Ile Gln Ile Asp Asp Ser Met Arg Glu Thr Tyr Leu Lys Lys     370 375 380 Phe Gly Thr Ser Ser Ala Gly His Ser Ser Ser Thr Ser Ala Val Ala 385 390 395 400 Ser      <210> 27 <211> 1560 <212> DNA <213> Burkholderia cepacia <220> <221> CDS (222) (1) .. (1560) <400> 27 atg gac act tct gtg cag aag aag aaa ctc ggt tta aag aat cgc tac 48 Met Asp Thr Ser Val Gln Lys Lys Lys Leu Gly Leu Lys Asn Arg Tyr 1 5 10 15 gca gcg atg acc cgc ggt ctt ggc tgg cag acc agc tac cag ccg atg 96 Ala Ala Met Thr Arg Gly Leu Gly Trp Gln Thr Ser Tyr Gln Pro Met             20 25 30 gag aaa gtg ttt ccg tac gac aag tac gaa ggc atc aag atc cac gat 144 Glu Lys Val Phe Pro Tyr Asp Lys Tyr Glu Gly Ile Lys Ile His Asp         35 40 45 tgg gat aaa tgg gaa gac ccc ttc cgc ctg acc atg gac gcc tac tgg 192 Trp Asp Lys Trp Glu Asp Pro Phe Arg Leu Thr Met Asp Ala Tyr Trp     50 55 60 aaa tat cag ggc gag aag gaa aaa aag ctt tac gcc gtc atc gac gct 240 Lys Tyr Gln Gly Glu Lys Glu Lys Lys Leu Tyr Ala Val Ile Asp Ala 65 70 75 80 ttc gcg cag aac aac ggg cag ttg agc att tcc gac gcg cga tat gtc 288 Phe Ala Gln Asn Asn Gly Gln Leu Ser Ile Ser Asp Ala Arg Tyr Val                 85 90 95 aac gca ctc aag gtg ttt atc cag ggt gtg aca ccg ttg gag tat atg 336 Asn Ala Leu Lys Val Phe Ile Gln Gly Val Thr Pro Leu Glu Tyr Met             100 105 110 gca cac cga ggt ttt gcc cac att ggt cgg cat ttt acg ggt gaa ggg 384 Ala His Arg Gly Phe Ala His Ile Gly Arg His Phe Thr Gly Glu Gly         115 120 125 gca cgt gtt gct tgc cag atg cag tcc atc gac gag ctg cgt cac ttc 432 Ala Arg Val Ala Cys Gln Met Gln Ser Ile Asp Glu Leu Arg His Phe     130 135 140 cag acc gaa atg cat gct ctc tcg cac tac aac aag tat ttt aac ggt 480 Gln Thr Glu Met His Ala Leu Ser His Tyr Asn Lys Tyr Phe Asn Gly 145 150 155 160 ctg cac aac tcc atc cat tgg tac gac cgg gtt tgg tat ttg tcg gtg 528 Leu His Asn Ser Ile His Trp Tyr Asp Arg Val Trp Tyr Leu Ser Val                 165 170 175 ccc aag tca ttt ttt gaa gac gcg gcc acc ggt gga ccg ttc gag ttt 576 Pro Lys Ser Phe Phe Glu Asp Ala Ala Thr Gly Gly Pro Phe Glu Phe             180 185 190 ctt acc gcg gtg agc ttt tcg ttc gaa tat gtg ttg acc aac ctg ctg 624 Leu Thr Ala Val Ser Phe Ser Phe Glu Tyr Val Leu Thr Asn Leu Leu         195 200 205 ttt gtc ccc ttc atg tcg ggt gct gct tac aac ggg gac atg tct acg 672 Phe Val Pro Phe Met Ser Gly Ala Ala Tyr Asn Gly Asp Met Ser Thr     210 215 220 gtc act ttc ggt ttt tcg gcg caa agt gac gaa tcg cgc cac atg aca 720 Val Thr Phe Gly Phe Ser Ala Gln Ser Asp Glu Ser Arg His Met Thr 225 230 235 240 ctc ggc atc gaa tgc atc aag ttc atg cta gaa cag gat ccg gac aac 768 Leu Gly Ile Glu Cys Ile Lys Phe Met Leu Glu Gln Asp Pro Asp Asn                 245 250 255 gtg ccc atc gtg cag cgc tgg atc gac aag tgg ttc tgg cgc ggc tat 816 Val Pro Ile Val Gln Arg Trp Ile Asp Lys Trp Phe Trp Arg Gly Tyr             260 265 270 cgg ctg ttg agc atc gtg gcc atg atg cag gac tac atg ctg ccc aac 864 Arg Leu Leu Ser Ile Val Ala Met Met Gln Asp Tyr Met Leu Pro Asn         275 280 285 cgg gtg atg agc tgg cgc gag agc tgg gag atg tac gtc gag cag aac 912 Arg Val Met Ser Trp Arg Glu Ser Trp Glu Met Tyr Val Glu Gln Asn     290 295 300 ggc ggc gcg ctg ttc aag gat ctt gcg cgt tat ggc atc cgc aag ccc 960 Gly Gly Ala Leu Phe Lys Asp Leu Ala Arg Tyr Gly Ile Arg Lys Pro 305 310 315 320 aag ggc tgg gac cag gct tgc gaa ggc aag gac cac atc agc cat cag 1008 Lys Gly Trp Asp Gln Ala Cys Glu Gly Lys Asp His Ile Ser His Gln                 325 330 335 acc ttc gcc gta ttc tat aac tat aac gcc gcg gcc ccc atc cac acc 1056 Thr Phe Ala Val Phe Tyr Asn Tyr Asn Ala Ala Ala Pro Ile His Thr             340 345 350 tgg gtt ccc aca aaa gaa gaa atg gga tgg ctg tcg gag aag tac ccc 1104 Trp Val Pro Thr Lys Glu Glu Met Gly Trp Leu Ser Glu Lys Tyr Pro         355 360 365 gag acg ttc gac aag tat tac cgt ccg cgt tgg gac tac tgg cgc gag 1152 Glu Thr Phe Asp Lys Tyr Tyr Arg Pro Arg Trp Asp Tyr Trp Arg Glu     370 375 380 cag gcc gcc aag ggc aac cgt ttc tac aac aag acg ctg ccg atg ctc 1200 Gln Ala Ala Lys Gly Asn Arg Phe Tyr Asn Lys Thr Leu Pro Met Leu 385 390 395 400 tgc act acc tgc cag att ccg atg ata ttc acc gag cct ggc gac gca 1248 Cys Thr Thr Cys Gln Ile Pro Met Ile Phe Thr Glu Pro Gly Asp Ala                 405 410 415 acc aag atc tgc tat cgc gag tcg gcc tac ctc ggc gac aag tat cac 1296 Thr Lys Ile Cys Tyr Arg Glu Ser Ala Tyr Leu Gly Asp Lys Tyr His             420 425 430 ttc tgc agc gac cac tgc aag gag att ttt gac aac gaa ccc gaa aag 1344 Phe Cys Ser Asp His Cys Lys Glu Ile Phe Asp Asn Glu Pro Glu Lys         435 440 445 ttc gtg cag tca tgg ctt ccg ccg cag caa gtg tat caa gga aac tgt 1392 Phe Val Gln Ser Trp Leu Pro Pro Gln Gln Val Tyr Gln Gly Asn Cys     450 455 460 ttc aag ccg gat gcc gat ccg acc aag gag ggt ttt gat ccc ttg atg 1440 Phe Lys Pro Asp Ala Asp Pro Thr Lys Glu Gly Phe Asp Pro Leu Met 465 470 475 480 gcc ttg ctc gac tac tac aac ctg aat gta ggc cgg gac aac ttc gat 1488 Ala Leu Leu Asp Tyr Tyr Asn Leu Asn Val Gly Arg Asp Asn Phe Asp                 485 490 495 ttc gag gga tcg gaa gac caa aag aac ttt gct gcc tgg cgt gga gag 1536 Phe Glu Gly Ser Glu Asp Gln Lys Asn Phe Ala Ala Trp Arg Gly Glu             500 505 510 gtc ttg caa gga gaa gcc aaa tga 1560 Val Leu Gln Gly Glu Ala Lys         515 <210> 28 <211> 519 <212> PRT <213> Burkholderia cepacia <400> 28 Met Asp Thr Ser Val Gln Lys Lys Lys Leu Gly Leu Lys Asn Arg Tyr 1 5 10 15 Ala Ala Met Thr Arg Gly Leu Gly Trp Gln Thr Ser Tyr Gln Pro Met             20 25 30 Glu Lys Val Phe Pro Tyr Asp Lys Tyr Glu Gly Ile Lys Ile His Asp         35 40 45 Trp Asp Lys Trp Glu Asp Pro Phe Arg Leu Thr Met Asp Ala Tyr Trp     50 55 60 Lys Tyr Gln Gly Glu Lys Glu Lys Lys Leu Tyr Ala Val Ile Asp Ala 65 70 75 80 Phe Ala Gln Asn Asn Gly Gln Leu Ser Ile Ser Asp Ala Arg Tyr Val                 85 90 95 Asn Ala Leu Lys Val Phe Ile Gln Gly Val Thr Pro Leu Glu Tyr Met             100 105 110 Ala His Arg Gly Phe Ala His Ile Gly Arg His Phe Thr Gly Glu Gly         115 120 125 Ala Arg Val Ala Cys Gln Met Gln Ser Ile Asp Glu Leu Arg His Phe     130 135 140 Gln Thr Glu Met His Ala Leu Ser His Tyr Asn Lys Tyr Phe Asn Gly 145 150 155 160 Leu His Asn Ser Ile His Trp Tyr Asp Arg Val Trp Tyr Leu Ser Val                 165 170 175 Pro Lys Ser Phe Phe Glu Asp Ala Ala Thr Gly Gly Pro Phe Glu Phe             180 185 190 Leu Thr Ala Val Ser Phe Ser Phe Glu Tyr Val Leu Thr Asn Leu Leu         195 200 205 Phe Val Pro Phe Met Ser Gly Ala Ala Tyr Asn Gly Asp Met Ser Thr     210 215 220 Val Thr Phe Gly Phe Ser Ala Gln Ser Asp Glu Ser Arg His Met Thr 225 230 235 240 Leu Gly Ile Glu Cys Ile Lys Phe Met Leu Glu Gln Asp Pro Asp Asn                 245 250 255 Val Pro Ile Val Gln Arg Trp Ile Asp Lys Trp Phe Trp Arg Gly Tyr             260 265 270 Arg Leu Leu Ser Ile Val Ala Met Met Gln Asp Tyr Met Leu Pro Asn         275 280 285 Arg Val Met Ser Trp Arg Glu Ser Trp Glu Met Tyr Val Glu Gln Asn     290 295 300 Gly Gly Ala Leu Phe Lys Asp Leu Ala Arg Tyr Gly Ile Arg Lys Pro 305 310 315 320 Lys Gly Trp Asp Gln Ala Cys Glu Gly Lys Asp His Ile Ser His Gln                 325 330 335 Thr Phe Ala Val Phe Tyr Asn Tyr Asn Ala Ala Ala Pro Ile His Thr             340 345 350 Trp Val Pro Thr Lys Glu Glu Met Gly Trp Leu Ser Glu Lys Tyr Pro         355 360 365 Glu Thr Phe Asp Lys Tyr Tyr Arg Pro Arg Trp Asp Tyr Trp Arg Glu     370 375 380 Gln Ala Ala Lys Gly Asn Arg Phe Tyr Asn Lys Thr Leu Pro Met Leu 385 390 395 400 Cys Thr Thr Cys Gln Ile Pro Met Ile Phe Thr Glu Pro Gly Asp Ala                 405 410 415 Thr Lys Ile Cys Tyr Arg Glu Ser Ala Tyr Leu Gly Asp Lys Tyr His             420 425 430 Phe Cys Ser Asp His Cys Lys Glu Ile Phe Asp Asn Glu Pro Glu Lys         435 440 445 Phe Val Gln Ser Trp Leu Pro Pro Gln Gln Val Tyr Gln Gly Asn Cys     450 455 460 Phe Lys Pro Asp Ala Asp Pro Thr Lys Glu Gly Phe Asp Pro Leu Met 465 470 475 480 Ala Leu Leu Asp Tyr Tyr Asn Leu Asn Val Gly Arg Asp Asn Phe Asp                 485 490 495 Phe Glu Gly Ser Glu Asp Gln Lys Asn Phe Ala Ala Trp Arg Gly Glu             500 505 510 Val Leu Gln Gly Glu Ala Lys         515 <210> 29 <211> 996 <212> DNA <213> Burkholderia cepacia <220> <221> CDS (222) (1) .. (996) <400> 29 atg acc atc gat ttg aag acg cgg gaa atc aaa cca ctg cgt cac acc 48 Met Thr Ile Asp Leu Lys Thr Arg Glu Ile Lys Pro Leu Arg His Thr 1 5 10 15 tac acg cac gtg gct caa tac atc ggg gcc gat aaa gcc gct tcg cgc 96 Tyr Thr His Val Ala Gln Tyr Ile Gly Ala Asp Lys Ala Ala Ser Arg             20 25 30 tat cag gaa ggc act gta ggt gct caa ccc gca gcg aat ttt cat tac 144 Tyr Gln Glu Gly Thr Val Gly Ala Gln Pro Ala Ala Asn Phe His Tyr         35 40 45 cgg ccc acg tgg gat ccc gag cat gaa ctg ttc gac acg tcg cgt acc 192 Arg Pro Thr Trp Asp Pro Glu His Glu Leu Phe Asp Thr Ser Arg Thr     50 55 60 gcg att caa atg aag gac tgg tat gcg ctg aaa gac ccg cgt cag ttc 240 Ala Ile Gln Met Lys Asp Trp Tyr Ala Leu Lys Asp Pro Arg Gln Phe 65 70 75 80 tac tac gcg tcg tgg acg atg acc cga gcg cgg cag caa gac gcg atg 288 Tyr Tyr Ala Ser Trp Thr Met Thr Arg Ala Arg Gln Gln Asp Ala Met                 85 90 95 gaa tcc aac ttc gag ttt gtc gag tcg cgc ggc atg atc gat ctc gtt 336 Glu Ser Asn Phe Glu Phe Val Glu Ser Arg Gly Met Ile Asp Leu Val             100 105 110 tcc gat gag gtt cga caa cgg gcg ctt tcc gtt ctc gtg cct ttg cgt 384 Ser Asp Glu Val Arg Gln Arg Ala Leu Ser Val Leu Val Pro Leu Arg         115 120 125 cac gcg gcc tgg ggc gcg aac atg aac aac tcc cag atc tgt gcc cta 432 His Ala Ala Trp Gly Ala Asn Met Asn Asn Ser Gln Ile Cys Ala Leu     130 135 140 ggt tat ggc acg acc ttc act gcg ccg gct atg ttc cac gca atg gac 480 Gly Tyr Gly Thr Thr Phe Thr Ala Pro Ala Met Phe His Ala Met Asp 145 150 155 160 aat ctg ggt gta gcg cag tat ctc aca cga ctg gcg ctg gta atg tct 528 Asn Leu Gly Val Ala Gln Tyr Leu Thr Arg Leu Ala Leu Val Met Ser                 165 170 175 gga ccc gat ctt ctt gac gaa gcc aag caa gcc tgg atg acg agt ccc 576 Gly Pro Asp Leu Leu Asp Glu Ala Lys Gln Ala Trp Met Thr Ser Pro             180 185 190 gat tgg caa ccg ttg cgt cgt tat gtg gaa aac act ctg gtg ctg caa 624 Asp Trp Gln Pro Leu Arg Arg Tyr Val Glu Asn Thr Leu Val Leu Gln         195 200 205 gat ccg gtg gaa ctg ttc atc gcc caa aat ctg gcg ctc gac ggt ctt 672 Asp Pro Val Glu Leu Phe Ile Ala Gln Asn Leu Ala Leu Asp Gly Leu     210 215 220 ctt tat ccc atg atc tac ggc gct ttc gtc gac gat tac atc gca ctc 720 Leu Tyr Pro Met Ile Tyr Gly Ala Phe Val Asp Asp Tyr Ile Ala Leu 225 230 235 240 aac ggt ggt agc gca gtg gca atg cta acc act ttc atg ccc gag tgg 768 Asn Gly Gly Ser Ala Val Ala Met Leu Thr Thr Phe Met Pro Glu Trp                 245 250 255 cat gac gaa tcc agt cgc tgg gtc gat gcg gta gta aag acc atg gcg 816 His Asp Glu Ser Ser Arg Trp Val Asp Ala Val Val Lys Thr Met Ala             260 265 270 acg gaa tcg gag gat aac aaa gcg ctg ctc att cac tgg ttg cgt acc 864 Thr Glu Ser Glu Asp Asn Lys Ala Leu Leu Ile His Trp Leu Arg Thr         275 280 285 tgg gaa gat cag gcg gcg tca gcg ttg ttg cct gtc gct gaa atg gct 912 Trp Glu Asp Gln Ala Ala Ser Ala Leu Leu Pro Val Ala Glu Met Ala     290 295 300 ttg gcg gaa aac ggc cac gac gcc ttg gaa gaa gta agg cag caa ctt 960 Leu Ala Glu Asn Gly His Asp Ala Leu Glu Glu Val Arg Gln Gln Leu 305 310 315 320 cgt gcc cgc gtt gcg aag gcc ggg att gtt ctg taa 996 Arg Ala Arg Val Ala Lys Ala Gly Ile Val Leu                 325 330 <210> 30 <211> 331 <212> PRT <213> Burkholderia cepacia <400> 30 Met Thr Ile Asp Leu Lys Thr Arg Glu Ile Lys Pro Leu Arg His Thr 1 5 10 15 Tyr Thr His Val Ala Gln Tyr Ile Gly Ala Asp Lys Ala Ala Ser Arg             20 25 30 Tyr Gln Glu Gly Thr Val Gly Ala Gln Pro Ala Ala Asn Phe His Tyr         35 40 45 Arg Pro Thr Trp Asp Pro Glu His Glu Leu Phe Asp Thr Ser Arg Thr     50 55 60 Ala Ile Gln Met Lys Asp Trp Tyr Ala Leu Lys Asp Pro Arg Gln Phe 65 70 75 80 Tyr Tyr Ala Ser Trp Thr Met Thr Arg Ala Arg Gln Gln Asp Ala Met                 85 90 95 Glu Ser Asn Phe Glu Phe Val Glu Ser Arg Gly Met Ile Asp Leu Val             100 105 110 Ser Asp Glu Val Arg Gln Arg Ala Leu Ser Val Leu Val Pro Leu Arg         115 120 125 His Ala Ala Trp Gly Ala Asn Met Asn Asn Ser Gln Ile Cys Ala Leu     130 135 140 Gly Tyr Gly Thr Thr Phe Thr Ala Pro Ala Met Phe His Ala Met Asp 145 150 155 160 Asn Leu Gly Val Ala Gln Tyr Leu Thr Arg Leu Ala Leu Val Met Ser                 165 170 175 Gly Pro Asp Leu Leu Asp Glu Ala Lys Gln Ala Trp Met Thr Ser Pro             180 185 190 Asp Trp Gln Pro Leu Arg Arg Tyr Val Glu Asn Thr Leu Val Leu Gln         195 200 205 Asp Pro Val Glu Leu Phe Ile Ala Gln Asn Leu Ala Leu Asp Gly Leu     210 215 220 Leu Tyr Pro Met Ile Tyr Gly Ala Phe Val Asp Asp Tyr Ile Ala Leu 225 230 235 240 Asn Gly Gly Ser Ala Val Ala Met Leu Thr Thr Phe Met Pro Glu Trp                 245 250 255 His Asp Glu Ser Ser Arg Trp Val Asp Ala Val Val Lys Thr Met Ala             260 265 270 Thr Glu Ser Glu Asp Asn Lys Ala Leu Leu Ile His Trp Leu Arg Thr         275 280 285 Trp Glu Asp Gln Ala Ala Ser Ala Leu Leu Pro Val Ala Glu Met Ala     290 295 300 Leu Ala Glu Asn Gly His Asp Ala Leu Glu Glu Val Arg Gln Gln Leu 305 310 315 320 Arg Ala Arg Val Ala Lys Ala Gly Ile Val Leu                 325 330 <210> 31 <211> 357 <212> DNA <213> Burkholderia cepacia <220> <221> CDS (222) (1) .. (357) <400> 31 atg agc gtt gtt gcc ctc aaa ccc tac aag ttc ccg gca cga gac gcg 48 Met Ser Val Val Ala Leu Lys Pro Tyr Lys Phe Pro Ala Arg Asp Ala 1 5 10 15 cgc gaa aac ttt ccg gcg ccg ttg ctg ttt atc ggc tgg gaa gac cat 96 Arg Glu Asn Phe Pro Ala Pro Leu Leu Phe Ile Gly Trp Glu Asp His             20 25 30 ctg ttg ttt gcg gca cct gtt gcc ttg ccc ctg ccg tcg gac acg ttg 144 Leu Leu Phe Ala Ala Pro Val Ala Leu Pro Leu Pro Ser Asp Thr Leu         35 40 45 ttc ggt gcg ctg tgc acc cag gtg ttg ccc ggc act tat ggc tat cac 192 Phe Gly Ala Leu Cys Thr Gln Val Leu Pro Gly Thr Tyr Gly Tyr His     50 55 60 ccc gat ttc tca aag atc gac tgg agc cag gtg cag tgg ttt aag tcc 240 Pro Asp Phe Ser Lys Ile Asp Trp Ser Gln Val Gln Trp Phe Lys Ser 65 70 75 80 ggc cag ccg tgg cat ccc gac ccg gcg aag tcg ctg gct gaa aac ggt 288 Gly Gln Pro Trp His Pro Asp Pro Ala Lys Ser Leu Ala Glu Asn Gly                 85 90 95 ctg acg cac aaa gac gtg atc cgc ttt cgc acg cct ggc ttg aac ggt 336 Leu Thr His Lys Asp Val Ile Arg Phe Arg Thr Pro Gly Leu Asn Gly             100 105 110 ctg agc ggt tcc tgc aat tga 357 Leu Ser Gly Ser Cys Asn         115 <210> 32 <211> 118 <212> PRT <213> Burkholderia cepacia <400> 32 Met Ser Val Val Ala Leu Lys Pro Tyr Lys Phe Pro Ala Arg Asp Ala 1 5 10 15 Arg Glu Asn Phe Pro Ala Pro Leu Leu Phe Ile Gly Trp Glu Asp His             20 25 30 Leu Leu Phe Ala Ala Pro Val Ala Leu Pro Leu Pro Ser Asp Thr Leu         35 40 45 Phe Gly Ala Leu Cys Thr Gln Val Leu Pro Gly Thr Tyr Gly Tyr His     50 55 60 Pro Asp Phe Ser Lys Ile Asp Trp Ser Gln Val Gln Trp Phe Lys Ser 65 70 75 80 Gly Gln Pro Trp His Pro Asp Pro Ala Lys Ser Leu Ala Glu Asn Gly                 85 90 95 Leu Thr His Lys Asp Val Ile Arg Phe Arg Thr Pro Gly Leu Asn Gly             100 105 110 Leu Ser Gly Ser Cys Asn         115 <210> 33 <211> 1143 <212> DNA <213> Bacillus stearothermophilus <220> <221> CDS (222) (1) .. (1143) <400> 33 atg gaa aaa aat aaa atg tta ata gaa gaa aag ttg gac act gct gct 48 Met Glu Lys Asn Lys Met Leu Ile Glu Glu Lys Leu Asp Thr Ala Ala 1 5 10 15 ctt ctt gct aag gcg gag gaa ata ggc cgg att gct gag gaa gag gcg 96 Leu Leu Ala Lys Ala Glu Glu Ile Gly Arg Ile Ala Glu Glu Glu Ala             20 25 30 ggt gaa gcg gac cgc aat gcc tgt ttc tcc gac cgg gtg gct agg gcc 144 Gly Glu Ala Asp Arg Asn Ala Cys Phe Ser Asp Arg Val Ala Arg Ala         35 40 45 att aaa gaa gct gga ttc cac aag ctc atg cgt ccc aag cag tac gga 192 Ile Lys Glu Ala Gly Phe His Lys Leu Met Arg Pro Lys Gln Tyr Gly     50 55 60 gga ctg caa gta gac ttg cga act tac ggg gag att gtc cgc aca gtg 240 Gly Leu Gln Val Asp Leu Arg Thr Tyr Gly Glu Ile Val Arg Thr Val 65 70 75 80 gcc cgg tac agt gtt gcc gca gga tgg ctg acc tat ttt tat tcc atg 288 Ala Arg Tyr Ser Val Ala Ala Gly Trp Leu Thr Tyr Phe Tyr Ser Met                 85 90 95 cat gag gtt tgg gct gca tat ctg cct cca aaa ggc aga gaa gaa att 336 His Glu Val Trp Ala Ala Tyr Leu Pro Pro Lys Gly Arg Glu Glu Ile             100 105 110 ttt gga caa gga ggg ctg ttg gca gac gtc gtt gcc cct gtt ggc cgg 384 Phe Gly Gln Gly Gly Leu Leu Ala Asp Val Val Ala Pro Val Gly Arg         115 120 125 gtg gag aag gac ggg gac ggc tac cgt ctc tat ggg cag tgg aac ttc 432 Val Glu Lys Asp Gly Asp Gly Tyr Arg Leu Tyr Gly Gln Trp Asn Phe     130 135 140 tgt agc ggt gtc ctc cat agt gac tgg atc gga ctt ggc gcc atg atg 480 Cys Ser Gly Val Leu His Ser Asp Trp Ile Gly Leu Gly Ala Met Met 145 150 155 160 gag ctg cct gac ggc aat agt cct gag tac tgt ttg tta gtg ctg cct 528 Glu Leu Pro Asp Gly Asn Ser Pro Glu Tyr Cys Leu Leu Val Leu Pro                 165 170 175 aag tcg gat gtc cag atc gta gaa aat tgg gat acc atg ggc ctc cgc 576 Lys Ser Asp Val Gln Ile Val Glu Asn Trp Asp Thr Met Gly Leu Arg             180 185 190 gct tcg gga agc aac ggg gta tta gtt gaa ggt gct tat gtt cca tta 624 Ala Ser Gly Ser Asn Gly Val Leu Val Glu Gly Ala Tyr Val Pro Leu         195 200 205 cac cgg atc ttt ccg gct ggc cgg gtg atg gct cat ccg ctt ttc ttg 672 His Arg Ile Phe Pro Ala Gly Arg Val Met Ala His Pro Leu Phe Leu     210 215 220 ctt ggg ttc cct tta gta tct tta ggc ggc gac gaa cga ttg gtg tca 720 Leu Gly Phe Pro Leu Val Ser Leu Gly Gly Asp Glu Arg Leu Val Ser 225 230 235 240 ctt ttc caa gaa cgc act gag aag cgc att cgt gtc ttc aaa ggc ggc 768 Leu Phe Gln Glu Arg Thr Glu Lys Arg Ile Arg Val Phe Lys Gly Gly                 245 250 255 gcg aaa gaa aag gat tct gcc gct agc cag cgg ctg tta gcc gag atg 816 Ala Lys Glu Lys Asp Ser Ala Ala Ser Gln Arg Leu Leu Ala Glu Met             260 265 270 aaa aca gaa tta aat gca atg gaa ggc att gtg gaa caa tat atc cgc 864 Lys Thr Glu Leu Asn Ala Met Glu Gly Ile Val Glu Gln Tyr Ile Arg         275 280 285 cag ctt gag gct tgc caa aaa gaa gga aag acg gtg atg aac gat atg 912 Gln Leu Glu Ala Cys Gln Lys Glu Gly Lys Thr Val Met Asn Asp Met     290 295 300 gag cga gag cag cta ttc gca tgg cgt gga tat gtg gca aaa gcg tcc 960 Glu Arg Glu Gln Leu Phe Ala Trp Arg Gly Tyr Val Ala Lys Ala Ser 305 310 315 320 gcc aat att gcc gtc aga aca ctg tta act ctt gga ggc aat tcg atc 1008 Ala Asn Ile Ala Val Arg Thr Leu Leu Thr Leu Gly Gly Asn Ser Ile                 325 330 335 ttt aaa ggc gat ccg gta gaa ctg ttc aca aga gat ttg cta gcg gtg 1056 Phe Lys Gly Asp Pro Val Glu Leu Phe Thr Arg Asp Leu Leu Ala Val             340 345 350 gcc gca cat cct aac tcc ctg tgg gag gat gcg atg gct gca tat gga 1104 Ala Ala His Pro Asn Ser Leu Trp Glu Asp Ala Met Ala Ala Tyr Gly         355 360 365 aga acg ata ttc ggg ctg cca ggg gac cca gtc tgg taa 1143 Arg Thr Ile Phe Gly Leu Pro Gly Asp Pro Val Trp     370 375 380 <210> 34 <211> 380 <212> PRT <213> Bacillus stearothermophilus <400> 34 Met Glu Lys Asn Lys Met Leu Ile Glu Glu Lys Leu Asp Thr Ala Ala 1 5 10 15 Leu Leu Ala Lys Ala Glu Glu Ile Gly Arg Ile Ala Glu Glu Glu Ala             20 25 30 Gly Glu Ala Asp Arg Asn Ala Cys Phe Ser Asp Arg Val Ala Arg Ala         35 40 45 Ile Lys Glu Ala Gly Phe His Lys Leu Met Arg Pro Lys Gln Tyr Gly     50 55 60 Gly Leu Gln Val Asp Leu Arg Thr Tyr Gly Glu Ile Val Arg Thr Val 65 70 75 80 Ala Arg Tyr Ser Val Ala Ala Gly Trp Leu Thr Tyr Phe Tyr Ser Met                 85 90 95 His Glu Val Trp Ala Ala Tyr Leu Pro Pro Lys Gly Arg Glu Glu Ile             100 105 110 Phe Gly Gln Gly Gly Leu Leu Ala Asp Val Val Ala Pro Val Gly Arg         115 120 125 Val Glu Lys Asp Gly Asp Gly Tyr Arg Leu Tyr Gly Gln Trp Asn Phe     130 135 140 Cys Ser Gly Val Leu His Ser Asp Trp Ile Gly Leu Gly Ala Met Met 145 150 155 160 Glu Leu Pro Asp Gly Asn Ser Pro Glu Tyr Cys Leu Leu Val Leu Pro                 165 170 175 Lys Ser Asp Val Gln Ile Val Glu Asn Trp Asp Thr Met Gly Leu Arg             180 185 190 Ala Ser Gly Ser Asn Gly Val Leu Val Glu Gly Ala Tyr Val Pro Leu         195 200 205 His Arg Ile Phe Pro Ala Gly Arg Val Met Ala His Pro Leu Phe Leu     210 215 220 Leu Gly Phe Pro Leu Val Ser Leu Gly Gly Asp Glu Arg Leu Val Ser 225 230 235 240 Leu Phe Gln Glu Arg Thr Glu Lys Arg Ile Arg Val Phe Lys Gly Gly                 245 250 255 Ala Lys Glu Lys Asp Ser Ala Ala Ser Gln Arg Leu Leu Ala Glu Met             260 265 270 Lys Thr Glu Leu Asn Ala Met Glu Gly Ile Val Glu Gln Tyr Ile Arg         275 280 285 Gln Leu Glu Ala Cys Gln Lys Glu Gly Lys Thr Val Met Asn Asp Met     290 295 300 Glu Arg Glu Gln Leu Phe Ala Trp Arg Gly Tyr Val Ala Lys Ala Ser 305 310 315 320 Ala Asn Ile Ala Val Arg Thr Leu Leu Thr Leu Gly Gly Asn Ser Ile                 325 330 335 Phe Lys Gly Asp Pro Val Glu Leu Phe Thr Arg Asp Leu Leu Ala Val             340 345 350 Ala Ala His Pro Asn Ser Leu Trp Glu Asp Ala Met Ala Ala Tyr Gly         355 360 365 Arg Thr Ile Phe Gly Leu Pro Gly Asp Pro Val Trp     370 375 380 <210> 35 <211> 1191 <212> DNA <213> Helianthus annuus <220> <221> CDS (222) (1) .. (1191) <400> 35 atg gcg att cgc atc aat acg gcg acg ttt caa tca gac ctg tac cgt 48 Met Ala Ile Arg Ile Asn Thr Ala Thr Phe Gln Ser Asp Leu Tyr Arg 1 5 10 15 tca ttc gcg ttt cct caa ccg aaa cct ctc aga tct ccc aaa ttc gcc 96 Ser Phe Ala Phe Pro Gln Pro Lys Pro Leu Arg Ser Pro Lys Phe Ala             20 25 30 atg gct tcc acc att gga tcc gct aca acg aag gtt gaa agc acc aaa 144 Met Ala Ser Thr Ile Gly Ser Ala Thr Thr Lys Val Glu Ser Thr Lys         35 40 45 aag ccc ttt acc cct cca agg gag gtt cac caa cag gtg cta cac tca 192 Lys Pro Phe Thr Pro Pro Arg Glu Val His Gln Gln Val Leu His Ser     50 55 60 atg ccg cca caa aag atc gaa atc ttc aaa tcc atg gag ggt tgg gcc 240 Met Pro Pro Gln Lys Ile Glu Ile Phe Lys Ser Met Glu Gly Trp Ala 65 70 75 80 gaa aat aac ata ttg gtt cac cta aag cct gtc gaa aaa tgc tgg caa 288 Glu Asn Asn Ile Leu Val His Leu Lys Pro Val Glu Lys Cys Trp Gln                 85 90 95 gca cag gat ttc cta cca gat ccc gca tct gac gga ttt atg gaa caa 336 Ala Gln Asp Phe Leu Pro Asp Pro Ala Ser Asp Gly Phe Met Glu Gln             100 105 110 gtg gag gaa tta cgg gct cgg gct aag gag att ccg gat gat tac ttt 384 Val Glu Glu Leu Arg Ala Arg Ala Lys Glu Ile Pro Asp Asp Tyr Phe         115 120 125 gtt gtt ttg gtt gga gat atg att act gaa gaa gca ctg cct act tac 432 Val Val Leu Val Gly Asp Met Ile Thr Glu Glu Ala Leu Pro Thr Tyr     130 135 140 caa aca atg ctt aat act ctt gat ggt gtg cgt gat gag acc ggg gct 480 Gln Thr Met Leu Asn Thr Leu Asp Gly Val Arg Asp Glu Thr Gly Ala 145 150 155 160 acc cta ctt ctt ggg cta gtc tgg act cgg gct tgg acc gct gaa gaa 528 Thr Leu Leu Leu Gly Leu Val Trp Thr Arg Ala Trp Thr Ala Glu Glu                 165 170 175 aac agg cac ggt gat ctt cta cat cag tat ctg tat ctt agt ggg cgg 576 Asn Arg His Gly Asp Leu Leu His Gln Tyr Leu Tyr Leu Ser Gly Arg             180 185 190 gtc gac atg agg cag att cag aag aca att cag tac ctc att ggg tct 624 Val Asp Met Arg Gln Ile Gln Lys Thr Ile Gln Tyr Leu Ile Gly Ser         195 200 205 gga atg gac ccc cgg acc gaa aac agt cct tac ctt ggg ttc atc tac 672 Gly Met Asp Pro Arg Thr Glu Asn Ser Pro Tyr Leu Gly Phe Ile Tyr     210 215 220 act tca ttt caa gag cgt gcc acc ttc atc tct cac gga aac aca gcc 720 Thr Ser Phe Gln Glu Arg Ala Thr Phe Ile Ser His Gly Asn Thr Ala 225 230 235 240 cgg cac gca aag gag cat ggt gac gtg aag ctg gct caa atg tgc ggt 768 Arg His Ala Lys Glu His Gly Asp Val Lys Leu Ala Gln Met Cys Gly                 245 250 255 ata att gca gct gat gaa aaa agg cac gaa acc gcc tac aca aaa ata 816 Ile Ile Ala Ala Asp Glu Lys Arg His Glu Thr Ala Tyr Thr Lys Ile             260 265 270 gta gaa aaa ctc ttc gaa att gac ccg gac ggc act gtt ctc gct ttt 864 Val Glu Lys Leu Phe Glu Ile Asp Pro Asp Gly Thr Val Leu Ala Phe         275 280 285 gcc gac atg atg agg aaa aag atc tcc atg cct gca cac ttg atg tac 912 Ala Asp Met Met Arg Lys Lys Ile Ser Met Pro Ala His Leu Met Tyr     290 295 300 gat ggg cgt gat gat aac ctc ttc gaa aat ttc tca gct gtt gcc caa 960 Asp Gly Arg Asp Asp Asn Leu Phe Glu Asn Phe Ser Ala Val Ala Gln 305 310 315 320 agg ctc ggt gtg tac act gcg aag gac tat gca gac att ctg gag ttt 1008 Arg Leu Gly Val Tyr Thr Ala Lys Asp Tyr Ala Asp Ile Leu Glu Phe                 325 330 335 ctg gtg ggc cgg tgg aag gtg gcg gat tta acc ggg ctt tct ggt gaa 1056 Leu Val Gly Arg Trp Lys Val Ala Asp Leu Thr Gly Leu Ser Gly Glu             340 345 350 ggg cgt aaa gcc caa gac tat gtg tgc ggg ctg gcc cca aga atc aga 1104 Gly Arg Lys Ala Gln Asp Tyr Val Cys Gly Leu Ala Pro Arg Ile Arg         355 360 365 agg ctt gag gag agg aac tcg gca agg gcg aag gaa agt gtg aac gtt 1152 Arg Leu Glu Glu Arg Asn Ser Ala Arg Ala Lys Glu Ser Val Asn Val     370 375 380 ccg ttc agc tgg atc ttt gat aga gaa gtg aag ctc tga 1191 Pro Phe Ser Trp Ile Phe Asp Arg Glu Val Lys Leu 385 390 395 <210> 36 <211> 396 <212> PRT <213> Helianthus annuus <400> 36 Met Ala Ile Arg Ile Asn Thr Ala Thr Phe Gln Ser Asp Leu Tyr Arg 1 5 10 15 Ser Phe Ala Phe Pro Gln Pro Lys Pro Leu Arg Ser Pro Lys Phe Ala             20 25 30 Met Ala Ser Thr Ile Gly Ser Ala Thr Thr Lys Val Glu Ser Thr Lys         35 40 45 Lys Pro Phe Thr Pro Pro Arg Glu Val His Gln Gln Val Leu His Ser     50 55 60 Met Pro Pro Gln Lys Ile Glu Ile Phe Lys Ser Met Glu Gly Trp Ala 65 70 75 80 Glu Asn Asn Ile Leu Val His Leu Lys Pro Val Glu Lys Cys Trp Gln                 85 90 95 Ala Gln Asp Phe Leu Pro Asp Pro Ala Ser Asp Gly Phe Met Glu Gln             100 105 110 Val Glu Glu Leu Arg Ala Arg Ala Lys Glu Ile Pro Asp Asp Tyr Phe         115 120 125 Val Val Leu Val Gly Asp Met Ile Thr Glu Glu Ala Leu Pro Thr Tyr     130 135 140 Gln Thr Met Leu Asn Thr Leu Asp Gly Val Arg Asp Glu Thr Gly Ala 145 150 155 160 Thr Leu Leu Leu Gly Leu Val Trp Thr Arg Ala Trp Thr Ala Glu Glu                 165 170 175 Asn Arg His Gly Asp Leu Leu His Gln Tyr Leu Tyr Leu Ser Gly Arg             180 185 190 Val Asp Met Arg Gln Ile Gln Lys Thr Ile Gln Tyr Leu Ile Gly Ser         195 200 205 Gly Met Asp Pro Arg Thr Glu Asn Ser Pro Tyr Leu Gly Phe Ile Tyr     210 215 220 Thr Ser Phe Gln Glu Arg Ala Thr Phe Ile Ser His Gly Asn Thr Ala 225 230 235 240 Arg His Ala Lys Glu His Gly Asp Val Lys Leu Ala Gln Met Cys Gly                 245 250 255 Ile Ile Ala Ala Asp Glu Lys Arg His Glu Thr Ala Tyr Thr Lys Ile             260 265 270 Val Glu Lys Leu Phe Glu Ile Asp Pro Asp Gly Thr Val Leu Ala Phe         275 280 285 Ala Asp Met Met Arg Lys Lys Ile Ser Met Pro Ala His Leu Met Tyr     290 295 300 Asp Gly Arg Asp Asp Asn Leu Phe Glu Asn Phe Ser Ala Val Ala Gln 305 310 315 320 Arg Leu Gly Val Tyr Thr Ala Lys Asp Tyr Ala Asp Ile Leu Glu Phe                 325 330 335 Leu Val Gly Arg Trp Lys Val Ala Asp Leu Thr Gly Leu Ser Gly Glu             340 345 350 Gly Arg Lys Ala Gln Asp Tyr Val Cys Gly Leu Ala Pro Arg Ile Arg         355 360 365 Arg Leu Glu Glu Arg Asn Ser Ala Arg Ala Lys Glu Ser Val Asn Val     370 375 380 Pro Phe Ser Trp Ile Phe Asp Arg Glu Val Lys Leu 385 390 395

Claims (17)

Use of hydrogen peroxide as an oxidant, catalyzed by a monooxygenase enzyme, including the generation of hydrogen peroxide below the rate at which hydrogen peroxide is used in the reaction simultaneously with an oxidation reaction catalyzed by a monooxygenase enzyme and using hydrogen peroxide as the oxidant And oxidative damage of monooxygenase occurs at low levels. The method of claim 1, wherein the K _m of the monooxygenase enzyme for H ₂ O ₂ is at least 15 nM. The method of claim 1 or 2, wherein the monooxygenase enzyme is a P450 enzyme. The method according to claim 1, wherein the production rate of H ₂ O ₂ is 3 μg or less per mg of enzyme. The method of claim 1, wherein the concentration of H ₂ O ₂ is 1 mM or less throughout the reaction. 6. The method of claim 1, wherein the reaction lasts for at least 240 minutes. 7. 7. The method of claim 1, wherein H ₂ O ₂ is produced by an electrochemical reaction. The method of claim 1, wherein H ₂ O ₂ is produced by an enzymatic reaction. The method of claim 8, wherein the enzyme is glucose oxidase. The method according to claim 1, wherein H ₂ O ₂ is produced by H ₂ O ₂ precursors. The method of claim 10, wherein the H ₂ O ₂ precursor is perborate, percarbonate, or perphosphate. The method of claim 1, wherein the substrate oxidized by the monooxygenase enzyme is an alkane, aromatic compound, terpenoid compound, alkene or fatty acid. Use of an electrode to produce H ₂ O ₂ in carrying out the oxidation reaction as defined in claim 7. Use of an enzyme to produce H ₂ O ₂ in carrying out the oxidation reaction as defined in claim 8 or 9. Use of perborate, percarbonate or superphosphate to produce H ₂ O ₂ in the oxidation reaction defined in claim 10. In the presence of H ₂ O ₂ or hydroxyl radical isolated H ₂ O ₂ or hydroxyl radical, which controls the concentration of the mono-oxide dioxygenase is catalyzed by the enzyme, which comprises hydrogen peroxide performing oxidation using as an oxidizing agent, mono A method of carrying out an oxidation reaction catalyzed by an oxygenase enzyme, utilizing hydrogen peroxide as an oxidant and oxidative damage of monooxygenase occurs at low levels. The method of claim 16, wherein the sequestrant is EDTA.