KR102051018B1 - Method for producing ornithine decarboxylase mutants and their applications - Google Patents

Abstract

The present invention relates to a method for producing mutant strains of ornithine decarboxylase, the characteristics of the mutant strains, genes encoding the mutant strains of ornithine decarbonatease, and a method for producing putrescine using the same.
The present invention has the effect of increasing putrescine productivity or production efficiency, suppressing side reactions, and reducing the cost of putrescine purification.

Description

METHOD FOR PRODUCING ORNITHINE DECARBOXYLASE MUTANTS AND THEIR APPLICATIONS}

The present invention relates to the production of mutant strains of ornithine decarboxylase, the characteristics of the mutant strains, genes encoding the mutant strains of ornithine decarbonatease, and putrescine synthesis using them.

The present invention provides ornithine decarboxylase derived from Lactobacillus that is improved through protein engineering variations. In addition, the mutant strain of ornithine decarboxylase of the present invention is significantly reduced in the production of the catharine due to side reactions in the production of putrescine, thereby simplifying the putrescine separation process and thereby reducing the production cost.

Due to the continuous depletion of petroleum resources, research is being actively conducted to synthesize polymer precursors from biomaterials rather than petroleum resources. In particular, putrescine (putrescine; 1,4-diamino-butane) is a material that can be used as a polymer precursor, as well as many applications in the chemical industry. Examples are 4,6-nylon, pharmaceutical precursors, surfactants and the like. At present, more than 10,000 tons of putrescine is produced per year by petroleum resources, but there are difficulties in production economically and environmentally. This is the reason for the increase in oil prices and the toxic substances produced during production. In order to overcome such limitations, active research on putrescine synthesis through biotechnology has recently been conducted. Examples include the synthesis of putrescine from ornithine and the production of putrescine in large quantities using the bioconversion function of sugars from cells.

Ornithine decarboxylase uses PLP (pyridoxal-5'-phosphate) as a coenzyme, and cleaves the terminal carboxyl group of ornithine to synthesize putrescine. In the prior art, most of the putrescine decarboxylase derived from Escherichia coli was used to produce putrescine, and in order to increase putrescine production, the expression of enzymes in host cells was regulated or reduced using various promoters. To facilitate the release of tresins out of the cell, metabolic changes were made to alter other parts of the enzyme, such as overexpressing the back-transporter or blocking pathways that degrade putrescine. However, no studies have yet been reported to reduce the side reaction activity of ornithine decarboxylase (synthesized with 1,5-diamino-pentane) and maintain the original substrate activity.

Korean Patent Publication 10-2009-0107920

The problem with bioconversion from ornithine to putrescine is an enzyme side reaction. In the case of ornithine decarboxylase, in addition to ornithine, which is an original substrate, lysine is also used as a substrate to synthesize cardaber. These various substrate specificities are reported to be properties that do not disappear in the course of evolution.

However, this side reaction of ornithine decarboxylase impedes commercial use. Purification of putrescine is usually carried out by distillation. Putrescine (H ₂ N (CH ₂ ) ₄ NH ₂ ) and cardamom (H ₂ N (CH ₂ ) ₅ NH ₂ ) are only one carbon. Since only (-CH _2- ) is different and has the same functional group (diamine group), the structure is very similar. Therefore, conventionally, there is a problem in that a lot of energy is required to remove impurities called cardaber in the process of purifying putrescine.

In the present invention, in consideration of the purification process, commercial use of substrate specificity based on ornithine decarboxylase, whose structure has already been identified, in order to reduce side reactions from which cardaber is produced and to increase or maintain the production of putrescine, which is originally active. To increase in an easy direction.

The present invention has the effect of increasing putrescine productivity or production efficiency and suppressing side reactions. In particular, the present invention has the effect of inhibiting the synthesis of cardaberine, which is one of the side reactions, and has the effect of reducing the cost of putrescine purification.

In addition, the present invention, through the mass production of putrescine it is possible to utilize a variety of polymer precursors, pharmaceuticals, chemical additives and the like.

1 is a schematic diagram of putrescine synthesis using ornithine decarboxylase using ornithine as a substrate in the present invention. It also represents the cardabourne synthesis pathway, a side reaction of ornithine decarboxylase to be inhibited.
Figure 2 confirms the activity of ornithine decarboxylase derived from a variety of, showing the relative activity of the reactivity (or side reaction) when using ornithine as a substrate and lysine as a substrate. ODC_Lb is from Lactobacillus (inducible), ODC_Sc is from saccharomyces cerevisiae (inducible), ODC_Ec is from E. coli (constitutive), and ODC_Ef is from E. coli (inducible).
Figure 3 shows purified wild type ornithine decarboxylase derived from Lactobacillus, 696th alanine of wild type decarboxylase as glutamic acid (A696E), 702th valine as glycine (V702G), wild type The 713th alanine of the decarboxylase is leucine (A713L), the 696th alanine of the wild type decarboxylase and the 713th alanine is glutamic acid and leucine (A696E / A713L), the 702th valine and 713 of the wild type decarboxylase The first alanine is glycine and leucine (V702G / A713L), the wild type decarboxylase 696th alanine, the 702th valine and the 713th alanine are glutamic acid, glycine and leucine (A696E / V702G / A713L), the wild type decarbonate The mutant strain (E698D / A713L) in which the 698th glutamic acid of the enzyme was replaced with aspartic acid (E698D), and the 698th glutamic acid of the wild type decarboxylase and the 713th alanine replaced with aspartic acid and leucine (E698D / A713L). Using a graph comparing after the determination of (a) ornithine, specific activity and (B) lysine-specific activity, respectively.
Fig. 4 shows that the 698th glutamic acid of purified Lactobacillus-derived wild type ornithine decarboxylase and wild type ornithine decarboxylase is aspartic acid (E698D), and the 713th alanine of wild type decarboxylase is leucine (A713L). The kinetic coefficients of lysine and ornithine were compared using the mutant strains (E698D / A713L) substituted with aspartic acid and leucine for 698-glutamic acid and 713-alanine of wild-type ornithine decarboxylase.
5 is a view comparing the bioconversion reaction under various conditions. (A) Putrescine synthesis was quantified by the ornithine substrate, and wild-type ornithine decarboxylase derived from purified Lactobacillus was reacted in 0.37 M concentration buffer (see q). When the mutant substituted with leucine was reacted in 0.37 M concentration buffer (see ○), when wild type ornithine decarboxylase was reacted in 0.1 M concentration buffer (see ◆), the 713th alanine of wild type decarboxylase was The mutant strain substituted with leucine is a diagram when (0.1) reacted in a 0.1 M concentration buffer. (B) When lysine was used to quantify the synthesis, and when the purified Lactobacillus-derived wild type ornithine decarboxylase was reacted in 0.37 M concentration buffer (see q), the 713th alanine of wild type decarboxylase was When mutant substituted with leucine was reacted in 0.37 M concentration buffer (see ○), wild type ornithine decarboxylase was reacted in 0.1 M concentration buffer (see ◆), 713th alanine of wild type decarboxylase was leucine It is a figure when (refer to ◇) the mutant strain substituted by the reaction in 0.1 M concentration buffer.
Figure 6 shows the amino acid sequence of ornithine decarboxylase variant strain derived from Lactobacillus used in the present invention,
(a) is the wild type amino acid sequence (SEQ ID NO: 1),
(b) shows the amino acid sequence of A696E (SEQ ID NO: 1-696 alanine substituted with glutamic acid) (SEQ ID NO: 2),
(c) shows the amino acid sequence of V702G (SEQ ID NO: 1-702 valine with glycine) (SEQ ID NO: 3),
(d) shows amino acid sequence of A713L (SEQ ID NO: 1 to 713 alanine substituted with leucine) (SEQ ID NO: 4)
(e) is the combinatorial amino acid sequence of A696E and A713L (SEQ ID NO: 5),
(f) is the V702G and A713L combinatorial amino acid sequence (SEQ ID NO: 6),
(g) is the combinatorial amino acid sequence of A696E and V702G and A713L (SEQ ID NO: 7),
(h) is the amino acid sequence of E698D (SEQ ID NOs: 198-6 glutamic acid replaced with aspartic acid) (SEQ ID NO: 8),
(i) is the combinatorial amino acid sequence of E698D and A713L (SEQ ID NO: 9).
Figure 7 shows the DNA sequence of the mutant strain of ornithine decarboxylase detected in the present invention,
(a) is the wild type DNA sequence (SEQ ID NO: 10),
(b) shows the DNA sequence of A696E (SEQ ID NO: 11),
(c) shows the DNA sequence of V702G (SEQ ID NO: 12),
(d) shows the DNA sequence of A713L (SEQ ID NO: 13),
(e) shows the combinatorial DNA sequence of A696E and A713L (SEQ ID NO: 14),
(f) shows the combinatorial DNA sequence of V702G and A713L (SEQ ID NO: 15),
(g) is the combinatorial DNA sequence of A696E and V702G and A713L (SEQ ID NO: 16),
(h) is the DNA sequence of E698D (SEQ ID NO: 17),
(i) is the combinatorial DNA sequence of E698D and A713L (SEQ ID NO: 18).

The terms used in the present invention are commonly used in the biological bioconversion industry, and those skilled in the art will understand the meaning thereof, but the following description briefly describes:

(1) Ornithine decarboxylase means an enzyme that removes carboxyl groups from ornithine.

(2) PLP (pyridoxal-5'-phosphate) added to ornithine decarboxylase reaction is a coenzyme of an enzyme essential for the reaction.

(3) The cell extract means a microbial extract of the present invention in which ornithine decarboxylase is expressed.

(4) Whole-cell reaction refers to a reaction using whole cell without breaking down the cell containing the specific enzyme and using the cell contents or separating and purifying the enzyme.

(5) Site saturated mutagenesis refers to the introduction of a change in the designated nucleotide sequence at a given position in a gene.

(6) Saturation mutagenesis refers to the introduction of changes in various nucleotide sequences at designated positions of genes. Saturation mutation refers to inserting a mutation through PCR by inserting a NNK codon instead of a sequence to be mutated on a primer of a complementary sequence that binds to the template strand. In this case, N in the NNK codon means A, T, G, C of the nucleotides and K means T, G.

(7) A vector refers to a polynucleotide consisting of single-stranded, double-stranded, circular or ultra-stranded DNA or RNA, and may include components that are operably linked at a suitable distance to produce a recombinant protein. Such components may include replication origins, promoters, enhancers, 5-RNA leader sequences, ribosomal binding sites, nucleic acid cassettes, termination and polyadenylation sites, or selectable label formats, which may be used for specific applications. So one or more may be missing. The nucleic acid cassette may comprise restriction enzyme sites for insertion of the recombinant protein to be expressed. In a functional vector, the nucleic acid cassette contains a nucleic acid sequence to be expressed that includes a translation initiation and termination site, and may use a vector capable of inserting two kinds of cassettes into the vector, if necessary, and the functions mentioned above may be added. Can be sequenced. The gene inserted into the recombinant vector may be E. coli strain BW25113 (DE3), BL21 (DE3) for expression, but may vary depending on the type of the inserted vector. Such vectors and expression strains can be readily selected by putrescine developers.

(8) Specific activity refers to the activity per unit of pure protein from which impurities and other proteins have been removed through enzymatic purification. Usually, the amount of enzyme catalyzing 1 μmol substrate change per minute is 1 unit. It is expressed in units per mg.

(9) Kinetic parameters indicate the substrate affinity and substrate conversion capacity of enzymes using substrate solutions of different concentrations.

In the present invention, a variant of ornithine decarboxylase represented by any one of the following amino acid sequences is provided:

An amino acid sequence of which the 713th alanine is substituted with any of the hydrophobic amino acids in the amino acid sequence of SEQ ID 1; or

And amino acid sequence 698 where glutamic acid is substituted with aspartic acid and 713 alanine is substituted with any one of hydrophobic amino acids.

The hydrophobic amino acid means an amino acid of any one of phenylalanine, tryptophan, isoleucine, leucine, proline, methionine, valine, and alanine. Preferably, the hydrophobic amino acid is leucine, isoleucine or valine, more preferably leucine.

The amino acid sequence may have at least 95% homology, preferably at least 98% homology, more preferably at least 99% homology, within the range of not lowering the activity of ornithine decarboxylase. .

The present invention also provides a DNA encoding a variant of ornithine decarboxylase.

The DNA may be used without limitation as long as it corresponds to a DNA encoding an amino acid provided in the present invention, and includes a sequence obtained by optimizing the DNA, wherein the DNA is preferably a DNA sequence represented by SEQ ID NO: 13 or 18 Can be.

The present invention also provides a recombinant DNA vector containing the DNA, a host cell transformed with the recombinant DNA vector, and an extract of the host cell.

The vector and the host cell can be used without limitation so long as those skilled in the art can easily select.

The present invention provides a method for producing putrescine, using any one selected from the group consisting of a host cell transformed with the recombinant DNA vector and an extract of the host cell as a biocatalyst.

In the production method, ornithine can be used as a substrate. It also includes synthesizing ornithine using a substance that is a precursor of ornithine as a substrate and then producing putrescine therefrom. Synthesis of ornithine can be used without limitation so long as it can be easily selected by those skilled in the art.

In the above production method, a buffer may be used as necessary. The buffer can be used without limitation, known to those skilled in the art, for example, citric-sodium citrate buffer and the like can be used. In addition, the concentration of the buffer may be preferably 0.1 M to 1 M, more preferably 0.2 M to 1 M, even more preferably 0.3 M to 1 M. When the concentration in the above range is used, there is an effect of increasing the production of putrescine and suppressing the production of cardaber.

Example

Although the specific method of this invention is demonstrated in detail as an Example, the technical scope of this invention is not limited to these Examples.

Example 1.Comparison of Activity of Ornithine Decarboxylase from Various Origins

The reactivity of ornithine decarboxylase from four different species was compared. The four wild types compared were from Lactobacillus (inducible), saccharomyces cerevisiae (inducible), E. coli (constitutive) and E. coli (inducible). Is derived. As an abbreviation for this, ODC_Lb is expressed as Lactobacillus (inducible), ODC_Sc as saccharomyces cerevisiae (inducible), ODC_Ec as E.coli (constitutive), and ODC_Ef as E.coli (inducible). After inserting the corresponding gene into the pET24ma vector, the E. coli BL21 (DE3) was used to express the protein under 0.1 mM IPTG 18 ℃ conditions. Since the initial reaction rate was compared at 45 ℃ using 10% cell extract. Reactivity was compared with 4 mM ornithine and 4 mM lysine as a substrate, respectively. The result is shown in FIG. 2.

As shown in Fig. 2, Fig. 2 shows the activity of enzymes of various origins, specifically, ornithine decarboxylase activity which is an active product of putrescine using ornithine as a substrate, and lysine as a substrate. Showed lysine decarboxylase activity that produced cardaber. Looking at the results of Figure 2, the lysine decarboxylase activity was all similar, the ornithine decarboxylase activity was the most excellent ornithine decarboxylase derived from Lactobacillus. That is, ornithine decarboxylase derived from Lactobacillus has the lowest generation rate of side reactions compared to ornithine decarboxylase activity.

Example 2 Selection of Variation Target Part of Ornithine Decarboxylase

The crystal structure of the Lactobacillus ornithine decarboxylase of the present invention is known, and it is possible to predict the tunnel entering and exiting the substrate through structural analysis. In order to select functional residues to perform saturation variation among the predicted tunnel portions, multiple sequence alignments using sequence information of bioinformatics were performed. Residues that retain amino acid residues at specific positions in the protein structure are excluded as variant residues because they play a very important role in the structure and function of the protein and, in particular, are likely to play a direct role in catalysis.

Example 3 Saturation Variation and Functional Variation of Functional Residues of Ornithine Decarboxylase

Saturation mutations were performed using NNK codons for selected functional residues, followed by screening for variant strain libraries. All libraries were subjected to primary and secondary screening. The primary screening was performed through ornithine whole cell responses, and mutants with similar or faster activity were selected as changes in absorbance when compared to wild strains. Secondary screening was carried out via lysine whole cell response and was selected if the responsiveness to lysine was lower than that of wild strains for the primary strains selected. For selected mutants, intrinsic activity when ornithine or lysine was used as the substrate was measured.

Specifically, the wild and mutant strains of ornithine decarboxylase transformed into E. coli BL21 (DE3) were expressed using 50TG of inducer IPTG, followed by pure protein using Ni-NTA column. Purified. First, after protein expression, cells were disrupted with an acoustic wave crusher, and cell extracts were obtained after centrifugation. Cell extracts were added to a column equilibrated with 50 mM phosphate buffer (pH8.0) added with 300 mM sodium chloride, and then combined with nickel resin (resin) at 0 ° C. for 1 hour. Then, the protein that failed to bind to the resin was drained and other proteins that were not specifically bound were removed with Tris buffer containing 50 mM imidazole. Finally, only the desired protein was eluted with Tris buffer containing 250 mM imidazole. The eluted protein was desalted using a filtration column to remove imidazole, and finally only the active protein was obtained. The same amount of protein was measured using the Bradford protein quantification kit. After the reaction, the intrinsic activity was measured.

Intrinsic activity of wild-type and mutant strains was determined by high-performance liquid chromatography (HPLC) analysis. The reaction was performed at 50 ° C. for 30 minutes to 300 minutes to obtain the average of three experiments. The initial reaction rate when the conversion yield of 10% to 25% was shown was measured. A cation exchange column was used and the mobile phase consisted of 0.6 g / L citric acid, 4 g / L tartaric acid, 1.4 g / L ethyldiamine, 5% methanol 95% water. As the pH buffer solution, citric-sodium citrate buffer was used for pH 5.0. Intrinsic activity of the mutant and wild strains is shown in FIG. 3.

As shown in Figure 3, Figure 3 is a measure of the intrinsic activity against wild type, A696E, V702G, A713L, A696E / A713L, V702G / A713L, A696E / V702G / A713L, E698D, E698D / A713L, the functional residue (A696, V702, A713, E698, A696) are all located in the tunnel.

Figure 3 (A) is a comparison of the intrinsic activity when using ornithine as a substrate, A696E, V702G, A713L, A696E / A713L, V702G / A713L, A696E / V702G / A713L, E698D, E698D / A713L 19.9%, 4.3%, 89.4%, 12.8%, 4.9%, 0.1%, 75.6%, and 74.4% of the wild type intrinsic activity remained, indicating that intrinsic activity was reduced when ornithine was used as a substrate. .

Figure 3 (b) is a comparison of the intrinsic activity when using lysine as a substrate, A696E, V702G, A713L, A696E / A713L, V702G / A713L, A696E / V702G / A713L, E698D, E698D / A713L mutant strain wild type With respect to the intrinsic activity of 16.9%, 0.6%, 42.4%, 4.4%, 0.9%, 0.7%, 50.8%, 29.2% of the remaining activity was confirmed that side reactions are suppressed.

Example 4 Identification of Kinetic Coefficients for Functional Residues of Ornithine Decarboxylase

Among the mutants used in Example 3, kinetic parameters (kinetic parameters) in various concentration conditions in order to more closely compare the characteristics of the wild type A713L, E698D, E698D / A713L that can produce 70% ornithine with the wild type Was confirmed using ornithine and lysine, respectively.

Specifically, the kinetics coefficients of wild and mutant strains of ornithine decarbonatease after protein purification were confirmed using ornithine at a concentration of 0.2 mM to 6 mM, and the pH buffer solution was pH 5.0 citrate buffer (citric- sodium citrate buffer) was used. The reaction volume was run at 200 μl. Lysine concentrations from 0.45 mM to 140 mM were used to confirm the kinetic coefficient for lysine and the remaining reaction conditions were the same as for the ornithine reaction. The analysis was carried out through the HPLC analysis method specified above and was taken as the average of three experiments. Kinetic coefficients of mutant and wild strains are shown in FIG. 4.

As shown in FIG. 4, the k _cat value for ornithine of wild type and A713L was similar to about 39 s ⁻¹ , and the A713L mutant showed 2.16 times lower k _cat value for lysine than the wild type. k _cat / K _M values for the mutant lysine due to the decrease in k _cat value of the product was confirmed to be 1.93-fold decreased. In conclusion, it was confirmed that the A713L mutant is a mutant that can reduce activity by distinguishing ornithine and lysine. E698D and E698D / A713L mutants also reduced k _cat values for lysine by 2.08 and 2.59 times, respectively. As a result, it was confirmed that k _cat / K _M values of lysine of E698D and E698D / A713L mutants were decreased by 1.28 and 1.67 times.

Example 5. Mutant strain characterization of ornithine decarboxylase

In the present invention, in order to examine the effect of the variation of the A713L single amino acid substitution mutant of the ornithine decarboxylase having the highest ornithine intrinsic activity, a high concentration of 51.5 g / L (0.39 M) ornithine was used as a substrate. (See Fig. 5A) and 2.57 g / L (17.6 mM) lysine were used as substrates (see Fig. 5B). In order to set the appropriate reaction conditions in the reaction under the two substrate conditions, a buffer concentration for titrating the pH was performed under two conditions (0.1 M or 0.37 M).

Specifically, 0.1 mg of wild and mutant strains of lysine decarboxylase after protein purification were used for the reaction. 0.39 M ornithine or 17.6 mM lysine was used as the substrate and 0.1 or 0.37 M citric-sodium citrate buffer was used as the buffer (pH 5.0). 0.1 mM PLP coenzyme was used and the reaction was 50? The reaction volume was at 2 mL. The results are shown in FIG. 5.

5A is a case where 51.5 g / L (0.39 M) of ornithine is used as a substrate, and a 0.37 M buffer is used for the reaction of converting ornithine to putrescine ( After 4 hours, wild-type and A713L mutants produced 33.0 g / L and 31.6 g / L of putrescine, respectively. In addition, when a low concentration of buffer (0.1 M) was used (see ◆ and ◇ in FIG. 5A), wild-type and A713L mutants showed 20.2 g / L and 20.7 g / L of putrescine after 7 hours. Generated. The putrescine production capacity of wild type and A713L mutant strains was similar, and it was confirmed that the use of high concentration buffer (0.37 M) is beneficial for reactivity.

5 (b) shows a case where 2.57 g / L (17.6 mM) lysine was used as a substrate, and 0.37 M of buffer was used for the experiment for converting lysine into cardamom. And wild type and A713L mutant strains produced 0.03 g / L and 0.007 g / L of cadaveric after 4 hours, respectively. In addition, when 0.1 M buffer was used (see ◆ and ◇ of FIG. 5), side reactions in which cardaber was produced increased, and wild type and A713L mutants were 0.59 g / L and 0.38 g after 7 h, respectively. / L of cardinalgy was generated as a side reaction. As a result, it was confirmed that the mutant strain suppressed side reactions that produce cardaber, and this effect was more pronounced when using a high concentration of buffer (0.37 M).

<110> Seoul National University R & DB Foundation <120> METHOD FOR PRODUCING ORNITHINE DECARBOXYLASE MUTANTS AND THEIR          APPLICATIONS <130> Y17KP-004 <160> 18 <170> KoPatentIn 3.0 <210> 1 <211> 731 <212> PRT <213> Artificial Sequence <220> <223> ORNITHINE DECARBOXYLASE MUTANT <400> 1 Met Ser Ser Ser Leu Lys Ile Ala Ser Thr Gln Glu Ala Arg Gln Tyr   1 5 10 15 Phe Asp Thr Asp Arg Val Val Val Asp Ala Val Gly Ser Asp Phe Thr              20 25 30 Asp Val Gly Ala Val Ile Ala Met Asp Tyr Glu Thr Asp Val Ile Asp          35 40 45 Ala Ala Asp Ala Thr Lys Phe Gly Ile Pro Val Phe Ala Val Thr Lys      50 55 60 Asp Ala Gln Ala Ile Ser Ala Asp Glu Leu Lys Lys Ile Phe His Ile  65 70 75 80 Ile Asp Leu Glu Asn Lys Phe Asp Ala Thr Val Asn Ala Arg Glu Ile                  85 90 95 Glu Thr Ala Val Asn Asn Tyr Glu Asp Ser Ile Leu Pro Pro Phe Phe             100 105 110 Lys Ser Leu Lys Glu Tyr Val Ser Arg Gly Leu Ile Gln Phe Asp Cys         115 120 125 Pro Gly His Gln Gly Gly Gln Tyr Tyr Arg Lys His Pro Ala Gly Arg     130 135 140 Glu Phe Tyr Asp Phe Phe Gly Glu Thr Val Phe Arg Ala Asp Leu Cys 145 150 155 160 Asn Ala Asp Val Ala Leu Gly Asp Leu Leu Ile His Glu Gly Pro Ala                 165 170 175 Val Ala Ala Glu Lys His Ala Ala Arg Val Tyr Asn Ala Asp Lys Thr             180 185 190 Tyr Phe Val Leu Gly Gly Ser Ser Asn Ala Asn Asn Thr Val Thr Ser         195 200 205 Ala Leu Val Ser Asn Gly Asp Leu Val Leu Phe Asp Arg Asn Asn His     210 215 220 Lys Ser Val Tyr Asn Ser Ala Leu Ala Met Ala Gly Gly Arg Pro Val 225 230 235 240 Tyr Leu Gln Thr Asn Arg Asn Pro Tyr Gly Phe Ile Gly Gly Ile Tyr                 245 250 255 Asp Ser Asp Phe Asp Glu Lys Lys Ile Arg Glu Leu Ala Ala Lys Val             260 265 270 Asp Pro Glu Arg Ala Lys Trp Lys Arg Pro Phe Arg Leu Ala Val Ile         275 280 285 Gln Leu Gly Thr Tyr Asp Gly Thr Ile Tyr Asn Ala His Glu Val Val     290 295 300 Lys Arg Ile Gly His Leu Cys Asp Tyr Ile Glu Phe Asp Ser Ala Trp 305 310 315 320 Val Gly Tyr Glu Gln Phe Ile Pro Met Met Arg Asn Ser Ser Pro Leu                 325 330 335 Leu Ile Asp Asp Leu Gly Pro Glu Asp Pro Gly Ile Ile Val Val Gln             340 345 350 Ser Val His Lys Gln Gln Ala Gly Phe Ser Gln Thr Ser Gln Ile His         355 360 365 Lys Lys Asp Ser His Ile Lys Gly Gln Leu Arg Tyr Cys Asp His Lys     370 375 380 His Phe Asn Asn Ser Phe Asn Leu Phe Met Ser Thr Ser Pro Phe Tyr 385 390 395 400 Pro Met Tyr Ala Ala Leu Asp Val Asn Ala Ala Met Gln Glu Gly Glu                 405 410 415 Ala Gly Arg Lys Leu Trp His Asp Leu Leu Ile Thr Thr Ile Glu Ala             420 425 430 Arg Lys Lys Leu Ile Lys Ala Gly Ser Met Phe Arg Pro Phe Val Pro         435 440 445 Pro Val Val Asn Gly Lys Lys Trp Glu Asp Gly Asp Thr Glu Asp Met     450 455 460 Ala Asn Asn Ile Asp Tyr Trp Arg Phe Glu Lys Gly Ala Lys Trp His 465 470 475 480 Ala Tyr Glu Gly Tyr Gly Asp Asn Gln Tyr Tyr Val Asp Pro Asn Lys                 485 490 495 Phe Met Leu Thr Thr Pro Gly Ile Asn Pro Glu Thr Gly Asp Tyr Glu             500 505 510 Asp Phe Gly Val Pro Ala Thr Ile Val Ala Asn Tyr Leu Arg Asp His         515 520 525 Gly Ile Ile Pro Glu Lys Ser Asp Leu Asn Ser Ile Leu Phe Leu Met     530 535 540 Thr Pro Ala Glu Thr Pro Ala Lys Met Asn Asn Leu Ile Thr Gln Leu 545 550 555 560 Leu Gln Leu Gln Arg Leu Ile Glu Glu Asp Ala Pro Leu Lys Gln Val                 565 570 575 Leu Pro Ser Ile Tyr Ala Ala Asn Glu Glu Arg Tyr Asn Gly Tyr Thr             580 585 590 Ile Arg Glu Leu Cys Gln Glu Leu His Asp Phe Tyr Lys Asn Asn Asn         595 600 605 Thr Phe Thr Tyr Gln Lys Arg Leu Phe Leu Arg Glu Phe Phe Pro Glu     610 615 620 Gln Gly Met Leu Pro Tyr Glu Ala Arg Gln Glu Phe Ile Arg Asn His 625 630 635 640 Asn Lys Leu Val Pro Leu Asn Lys Ile Glu Gly Glu Ile Ala Leu Glu                 645 650 655 Gly Ala Leu Pro Tyr Pro Pro Gly Val Phe Cys Val Ala Pro Gly Glu             660 665 670 Lys Trp Ser Glu Thr Ala Val Lys Tyr Phe Thr Ile Leu Gln Asp Gly         675 680 685 Ile Asn Asn Phe Pro Gly Phe Ala Pro Glu Ile Gln Gly Val Tyr Phe     690 695 700 Lys Gln Glu Gly Asp Lys Val Val Ala Tyr Gly Glu Val Tyr Asp Ala 705 710 715 720 Glu Val Ala Lys Asn Asp Asp Arg Tyr Asn Asn                 725 730 <210> 2 <211> 731 <212> PRT <213> Artificial Sequence <220> <223> ORNITHINE DECARBOXYLASE MUTANT <400> 2 Met Ser Ser Ser Leu Lys Ile Ala Ser Thr Gln Glu Ala Arg Gln Tyr   1 5 10 15 Phe Asp Thr Asp Arg Val Val Val Asp Ala Val Gly Ser Asp Phe Thr              20 25 30 Asp Val Gly Ala Val Ile Ala Met Asp Tyr Glu Thr Asp Val Ile Asp          35 40 45 Ala Ala Asp Ala Thr Lys Phe Gly Ile Pro Val Phe Ala Val Thr Lys      50 55 60 Asp Ala Gln Ala Ile Ser Ala Asp Glu Leu Lys Lys Ile Phe His Ile  65 70 75 80 Ile Asp Leu Glu Asn Lys Phe Asp Ala Thr Val Asn Ala Arg Glu Ile                  85 90 95 Glu Thr Ala Val Asn Asn Tyr Glu Asp Ser Ile Leu Pro Pro Phe Phe             100 105 110 Lys Ser Leu Lys Glu Tyr Val Ser Arg Gly Leu Ile Gln Phe Asp Cys         115 120 125 Pro Gly His Gln Gly Gly Gln Tyr Tyr Arg Lys His Pro Ala Gly Arg     130 135 140 Glu Phe Tyr Asp Phe Phe Gly Glu Thr Val Phe Arg Ala Asp Leu Cys 145 150 155 160 Asn Ala Asp Val Ala Leu Gly Asp Leu Leu Ile His Glu Gly Pro Ala                 165 170 175 Val Ala Ala Glu Lys His Ala Ala Arg Val Tyr Asn Ala Asp Lys Thr             180 185 190 Tyr Phe Val Leu Gly Gly Ser Ser Asn Ala Asn Asn Thr Val Thr Ser         195 200 205 Ala Leu Val Ser Asn Gly Asp Leu Val Leu Phe Asp Arg Asn Asn His     210 215 220 Lys Ser Val Tyr Asn Ser Ala Leu Ala Met Ala Gly Gly Arg Pro Val 225 230 235 240 Tyr Leu Gln Thr Asn Arg Asn Pro Tyr Gly Phe Ile Gly Gly Ile Tyr                 245 250 255 Asp Ser Asp Phe Asp Glu Lys Lys Ile Arg Glu Leu Ala Ala Lys Val             260 265 270 Asp Pro Glu Arg Ala Lys Trp Lys Arg Pro Phe Arg Leu Ala Val Ile         275 280 285 Gln Leu Gly Thr Tyr Asp Gly Thr Ile Tyr Asn Ala His Glu Val Val     290 295 300 Lys Arg Ile Gly His Leu Cys Asp Tyr Ile Glu Phe Asp Ser Ala Trp 305 310 315 320 Val Gly Tyr Glu Gln Phe Ile Pro Met Met Arg Asn Ser Ser Pro Leu                 325 330 335 Leu Ile Asp Asp Leu Gly Pro Glu Asp Pro Gly Ile Ile Val Val Gln             340 345 350 Ser Val His Lys Gln Gln Ala Gly Phe Ser Gln Thr Ser Gln Ile His         355 360 365 Lys Lys Asp Ser His Ile Lys Gly Gln Leu Arg Tyr Cys Asp His Lys     370 375 380 His Phe Asn Asn Ser Phe Asn Leu Phe Met Ser Thr Ser Pro Phe Tyr 385 390 395 400 Pro Met Tyr Ala Ala Leu Asp Val Asn Ala Ala Met Gln Glu Gly Glu                 405 410 415 Ala Gly Arg Lys Leu Trp His Asp Leu Leu Ile Thr Thr Ile Glu Ala             420 425 430 Arg Lys Lys Leu Ile Lys Ala Gly Ser Met Phe Arg Pro Phe Val Pro         435 440 445 Pro Val Val Asn Gly Lys Lys Trp Glu Asp Gly Asp Thr Glu Asp Met     450 455 460 Ala Asn Asn Ile Asp Tyr Trp Arg Phe Glu Lys Gly Ala Lys Trp His 465 470 475 480 Ala Tyr Glu Gly Tyr Gly Asp Asn Gln Tyr Tyr Val Asp Pro Asn Lys                 485 490 495 Phe Met Leu Thr Thr Pro Gly Ile Asn Pro Glu Thr Gly Asp Tyr Glu             500 505 510 Asp Phe Gly Val Pro Ala Thr Ile Val Ala Asn Tyr Leu Arg Asp His         515 520 525 Gly Ile Ile Pro Glu Lys Ser Asp Leu Asn Ser Ile Leu Phe Leu Met     530 535 540 Thr Pro Ala Glu Thr Pro Ala Lys Met Asn Asn Leu Ile Thr Gln Leu 545 550 555 560 Leu Gln Leu Gln Arg Leu Ile Glu Glu Asp Ala Pro Leu Lys Gln Val                 565 570 575 Leu Pro Ser Ile Tyr Ala Ala Asn Glu Glu Arg Tyr Asn Gly Tyr Thr             580 585 590 Ile Arg Glu Leu Cys Gln Glu Leu His Asp Phe Tyr Lys Asn Asn Asn         595 600 605 Thr Phe Thr Tyr Gln Lys Arg Leu Phe Leu Arg Glu Phe Phe Pro Glu     610 615 620 Gln Gly Met Leu Pro Tyr Glu Ala Arg Gln Glu Phe Ile Arg Asn His 625 630 635 640 Asn Lys Leu Val Pro Leu Asn Lys Ile Glu Gly Glu Ile Ala Leu Glu                 645 650 655 Gly Ala Leu Pro Tyr Pro Pro Gly Val Phe Cys Val Ala Pro Gly Glu             660 665 670 Lys Trp Ser Glu Thr Ala Val Lys Tyr Phe Thr Ile Leu Gln Asp Gly         675 680 685 Ile Asn Asn Phe Pro Gly Phe Glu Pro Glu Ile Gln Gly Val Tyr Phe     690 695 700 Lys Gln Glu Gly Asp Lys Val Val Ala Tyr Gly Glu Val Tyr Asp Ala 705 710 715 720 Glu Val Ala Lys Asn Asp Asp Arg Tyr Asn Asn                 725 730 <210> 3 <211> 731 <212> PRT <213> Artificial Sequence <220> <223> ORNITHINE DECARBOXYLASE MUTANT <400> 3 Met Ser Ser Ser Leu Lys Ile Ala Ser Thr Gln Glu Ala Arg Gln Tyr   1 5 10 15 Phe Asp Thr Asp Arg Val Val Val Asp Ala Val Gly Ser Asp Phe Thr              20 25 30 Asp Val Gly Ala Val Ile Ala Met Asp Tyr Glu Thr Asp Val Ile Asp          35 40 45 Ala Ala Asp Ala Thr Lys Phe Gly Ile Pro Val Phe Ala Val Thr Lys      50 55 60 Asp Ala Gln Ala Ile Ser Ala Asp Glu Leu Lys Lys Ile Phe His Ile  65 70 75 80 Ile Asp Leu Glu Asn Lys Phe Asp Ala Thr Val Asn Ala Arg Glu Ile                  85 90 95 Glu Thr Ala Val Asn Asn Tyr Glu Asp Ser Ile Leu Pro Pro Phe Phe             100 105 110 Lys Ser Leu Lys Glu Tyr Val Ser Arg Gly Leu Ile Gln Phe Asp Cys         115 120 125 Pro Gly His Gln Gly Gly Gln Tyr Tyr Arg Lys His Pro Ala Gly Arg     130 135 140 Glu Phe Tyr Asp Phe Phe Gly Glu Thr Val Phe Arg Ala Asp Leu Cys 145 150 155 160 Asn Ala Asp Val Ala Leu Gly Asp Leu Leu Ile His Glu Gly Pro Ala                 165 170 175 Val Ala Ala Glu Lys His Ala Ala Arg Val Tyr Asn Ala Asp Lys Thr             180 185 190 Tyr Phe Val Leu Gly Gly Ser Ser Asn Ala Asn Asn Thr Val Thr Ser         195 200 205 Ala Leu Val Ser Asn Gly Asp Leu Val Leu Phe Asp Arg Asn Asn His     210 215 220 Lys Ser Val Tyr Asn Ser Ala Leu Ala Met Ala Gly Gly Arg Pro Val 225 230 235 240 Tyr Leu Gln Thr Asn Arg Asn Pro Tyr Gly Phe Ile Gly Gly Ile Tyr                 245 250 255 Asp Ser Asp Phe Asp Glu Lys Lys Ile Arg Glu Leu Ala Ala Lys Val             260 265 270 Asp Pro Glu Arg Ala Lys Trp Lys Arg Pro Phe Arg Leu Ala Val Ile         275 280 285 Gln Leu Gly Thr Tyr Asp Gly Thr Ile Tyr Asn Ala His Glu Val Val     290 295 300 Lys Arg Ile Gly His Leu Cys Asp Tyr Ile Glu Phe Asp Ser Ala Trp 305 310 315 320 Val Gly Tyr Glu Gln Phe Ile Pro Met Met Arg Asn Ser Ser Pro Leu                 325 330 335 Leu Ile Asp Asp Leu Gly Pro Glu Asp Pro Gly Ile Ile Val Val Gln             340 345 350 Ser Val His Lys Gln Gln Ala Gly Phe Ser Gln Thr Ser Gln Ile His         355 360 365 Lys Lys Asp Ser His Ile Lys Gly Gln Leu Arg Tyr Cys Asp His Lys     370 375 380 His Phe Asn Asn Ser Phe Asn Leu Phe Met Ser Thr Ser Pro Phe Tyr 385 390 395 400 Pro Met Tyr Ala Ala Leu Asp Val Asn Ala Ala Met Gln Glu Gly Glu                 405 410 415 Ala Gly Arg Lys Leu Trp His Asp Leu Leu Ile Thr Thr Ile Glu Ala             420 425 430 Arg Lys Lys Leu Ile Lys Ala Gly Ser Met Phe Arg Pro Phe Val Pro         435 440 445 Pro Val Val Asn Gly Lys Lys Trp Glu Asp Gly Asp Thr Glu Asp Met     450 455 460 Ala Asn Asn Ile Asp Tyr Trp Arg Phe Glu Lys Gly Ala Lys Trp His 465 470 475 480 Ala Tyr Glu Gly Tyr Gly Asp Asn Gln Tyr Tyr Val Asp Pro Asn Lys                 485 490 495 Phe Met Leu Thr Thr Pro Gly Ile Asn Pro Glu Thr Gly Asp Tyr Glu             500 505 510 Asp Phe Gly Val Pro Ala Thr Ile Val Ala Asn Tyr Leu Arg Asp His         515 520 525 Gly Ile Ile Pro Glu Lys Ser Asp Leu Asn Ser Ile Leu Phe Leu Met     530 535 540 Thr Pro Ala Glu Thr Pro Ala Lys Met Asn Asn Leu Ile Thr Gln Leu 545 550 555 560 Leu Gln Leu Gln Arg Leu Ile Glu Glu Asp Ala Pro Leu Lys Gln Val                 565 570 575 Leu Pro Ser Ile Tyr Ala Ala Asn Glu Glu Arg Tyr Asn Gly Tyr Thr             580 585 590 Ile Arg Glu Leu Cys Gln Glu Leu His Asp Phe Tyr Lys Asn Asn Asn         595 600 605 Thr Phe Thr Tyr Gln Lys Arg Leu Phe Leu Arg Glu Phe Phe Pro Glu     610 615 620 Gln Gly Met Leu Pro Tyr Glu Ala Arg Gln Glu Phe Ile Arg Asn His 625 630 635 640 Asn Lys Leu Val Pro Leu Asn Lys Ile Glu Gly Glu Ile Ala Leu Glu                 645 650 655 Gly Ala Leu Pro Tyr Pro Pro Gly Val Phe Cys Val Ala Pro Gly Glu             660 665 670 Lys Trp Ser Glu Thr Ala Val Lys Tyr Phe Thr Ile Leu Gln Asp Gly         675 680 685 Ile Asn Asn Phe Pro Gly Phe Ala Pro Glu Ile Gln Gly Gly Tyr Phe     690 695 700 Lys Gln Glu Gly Asp Lys Val Val Ala Tyr Gly Glu Val Tyr Asp Ala 705 710 715 720 Glu Val Ala Lys Asn Asp Asp Arg Tyr Asn Asn                 725 730 <210> 4 <211> 731 <212> PRT <213> Artificial Sequence <220> <223> ORNITHINE DECARBOXYLASE MUTANT <400> 4 Met Ser Ser Ser Leu Lys Ile Ala Ser Thr Gln Glu Ala Arg Gln Tyr   1 5 10 15 Phe Asp Thr Asp Arg Val Val Val Asp Ala Val Gly Ser Asp Phe Thr              20 25 30 Asp Val Gly Ala Val Ile Ala Met Asp Tyr Glu Thr Asp Val Ile Asp          35 40 45 Ala Ala Asp Ala Thr Lys Phe Gly Ile Pro Val Phe Ala Val Thr Lys      50 55 60 Asp Ala Gln Ala Ile Ser Ala Asp Glu Leu Lys Lys Ile Phe His Ile  65 70 75 80 Ile Asp Leu Glu Asn Lys Phe Asp Ala Thr Val Asn Ala Arg Glu Ile                  85 90 95 Glu Thr Ala Val Asn Asn Tyr Glu Asp Ser Ile Leu Pro Pro Phe Phe             100 105 110 Lys Ser Leu Lys Glu Tyr Val Ser Arg Gly Leu Ile Gln Phe Asp Cys         115 120 125 Pro Gly His Gln Gly Gly Gln Tyr Tyr Arg Lys His Pro Ala Gly Arg     130 135 140 Glu Phe Tyr Asp Phe Phe Gly Glu Thr Val Phe Arg Ala Asp Leu Cys 145 150 155 160 Asn Ala Asp Val Ala Leu Gly Asp Leu Leu Ile His Glu Gly Pro Ala                 165 170 175 Val Ala Ala Glu Lys His Ala Ala Arg Val Tyr Asn Ala Asp Lys Thr             180 185 190 Tyr Phe Val Leu Gly Gly Ser Ser Asn Ala Asn Asn Thr Val Thr Ser         195 200 205 Ala Leu Val Ser Asn Gly Asp Leu Val Leu Phe Asp Arg Asn Asn His     210 215 220 Lys Ser Val Tyr Asn Ser Ala Leu Ala Met Ala Gly Gly Arg Pro Val 225 230 235 240 Tyr Leu Gln Thr Asn Arg Asn Pro Tyr Gly Phe Ile Gly Gly Ile Tyr                 245 250 255 Asp Ser Asp Phe Asp Glu Lys Lys Ile Arg Glu Leu Ala Ala Lys Val             260 265 270 Asp Pro Glu Arg Ala Lys Trp Lys Arg Pro Phe Arg Leu Ala Val Ile         275 280 285 Gln Leu Gly Thr Tyr Asp Gly Thr Ile Tyr Asn Ala His Glu Val Val     290 295 300 Lys Arg Ile Gly His Leu Cys Asp Tyr Ile Glu Phe Asp Ser Ala Trp 305 310 315 320 Val Gly Tyr Glu Gln Phe Ile Pro Met Met Arg Asn Ser Ser Pro Leu                 325 330 335 Leu Ile Asp Asp Leu Gly Pro Glu Asp Pro Gly Ile Ile Val Val Gln             340 345 350 Ser Val His Lys Gln Gln Ala Gly Phe Ser Gln Thr Ser Gln Ile His         355 360 365 Lys Lys Asp Ser His Ile Lys Gly Gln Leu Arg Tyr Cys Asp His Lys     370 375 380 His Phe Asn Asn Ser Phe Asn Leu Phe Met Ser Thr Ser Pro Phe Tyr 385 390 395 400 Pro Met Tyr Ala Ala Leu Asp Val Asn Ala Ala Met Gln Glu Gly Glu                 405 410 415 Ala Gly Arg Lys Leu Trp His Asp Leu Leu Ile Thr Thr Ile Glu Ala             420 425 430 Arg Lys Lys Leu Ile Lys Ala Gly Ser Met Phe Arg Pro Phe Val Pro         435 440 445 Pro Val Val Asn Gly Lys Lys Trp Glu Asp Gly Asp Thr Glu Asp Met     450 455 460 Ala Asn Asn Ile Asp Tyr Trp Arg Phe Glu Lys Gly Ala Lys Trp His 465 470 475 480 Ala Tyr Glu Gly Tyr Gly Asp Asn Gln Tyr Tyr Val Asp Pro Asn Lys                 485 490 495 Phe Met Leu Thr Thr Pro Gly Ile Asn Pro Glu Thr Gly Asp Tyr Glu             500 505 510 Asp Phe Gly Val Pro Ala Thr Ile Val Ala Asn Tyr Leu Arg Asp His         515 520 525 Gly Ile Ile Pro Glu Lys Ser Asp Leu Asn Ser Ile Leu Phe Leu Met     530 535 540 Thr Pro Ala Glu Thr Pro Ala Lys Met Asn Asn Leu Ile Thr Gln Leu 545 550 555 560 Leu Gln Leu Gln Arg Leu Ile Glu Glu Asp Ala Pro Leu Lys Gln Val                 565 570 575 Leu Pro Ser Ile Tyr Ala Ala Asn Glu Glu Arg Tyr Asn Gly Tyr Thr             580 585 590 Ile Arg Glu Leu Cys Gln Glu Leu His Asp Phe Tyr Lys Asn Asn Asn         595 600 605 Thr Phe Thr Tyr Gln Lys Arg Leu Phe Leu Arg Glu Phe Phe Pro Glu     610 615 620 Gln Gly Met Leu Pro Tyr Glu Ala Arg Gln Glu Phe Ile Arg Asn His 625 630 635 640 Asn Lys Leu Val Pro Leu Asn Lys Ile Glu Gly Glu Ile Ala Leu Glu                 645 650 655 Gly Ala Leu Pro Tyr Pro Pro Gly Val Phe Cys Val Ala Pro Gly Glu             660 665 670 Lys Trp Ser Glu Thr Ala Val Lys Tyr Phe Thr Ile Leu Gln Asp Gly         675 680 685 Ile Asn Asn Phe Pro Gly Phe Ala Pro Glu Ile Gln Gly Val Tyr Phe     690 695 700 Lys Gln Glu Gly Asp Lys Val Val Leu Tyr Gly Glu Val Tyr Asp Ala 705 710 715 720 Glu Val Ala Lys Asn Asp Asp Arg Tyr Asn Asn                 725 730 <210> 5 <211> 731 <212> PRT <213> Artificial Sequence <220> <223> ORNITHINE DECARBOXYLASE MUTANT <400> 5 Met Ser Ser Ser Leu Lys Ile Ala Ser Thr Gln Glu Ala Arg Gln Tyr   1 5 10 15 Phe Asp Thr Asp Arg Val Val Val Asp Ala Val Gly Ser Asp Phe Thr              20 25 30 Asp Val Gly Ala Val Ile Ala Met Asp Tyr Glu Thr Asp Val Ile Asp          35 40 45 Ala Ala Asp Ala Thr Lys Phe Gly Ile Pro Val Phe Ala Val Thr Lys      50 55 60 Asp Ala Gln Ala Ile Ser Ala Asp Glu Leu Lys Lys Ile Phe His Ile  65 70 75 80 Ile Asp Leu Glu Asn Lys Phe Asp Ala Thr Val Asn Ala Arg Glu Ile                  85 90 95 Glu Thr Ala Val Asn Asn Tyr Glu Asp Ser Ile Leu Pro Pro Phe Phe             100 105 110 Lys Ser Leu Lys Glu Tyr Val Ser Arg Gly Leu Ile Gln Phe Asp Cys         115 120 125 Pro Gly His Gln Gly Gly Gln Tyr Tyr Arg Lys His Pro Ala Gly Arg     130 135 140 Glu Phe Tyr Asp Phe Phe Gly Glu Thr Val Phe Arg Ala Asp Leu Cys 145 150 155 160 Asn Ala Asp Val Ala Leu Gly Asp Leu Leu Ile His Glu Gly Pro Ala                 165 170 175 Val Ala Ala Glu Lys His Ala Ala Arg Val Tyr Asn Ala Asp Lys Thr             180 185 190 Tyr Phe Val Leu Gly Gly Ser Ser Asn Ala Asn Asn Thr Val Thr Ser         195 200 205 Ala Leu Val Ser Asn Gly Asp Leu Val Leu Phe Asp Arg Asn Asn His     210 215 220 Lys Ser Val Tyr Asn Ser Ala Leu Ala Met Ala Gly Gly Arg Pro Val 225 230 235 240 Tyr Leu Gln Thr Asn Arg Asn Pro Tyr Gly Phe Ile Gly Gly Ile Tyr                 245 250 255 Asp Ser Asp Phe Asp Glu Lys Lys Ile Arg Glu Leu Ala Ala Lys Val             260 265 270 Asp Pro Glu Arg Ala Lys Trp Lys Arg Pro Phe Arg Leu Ala Val Ile         275 280 285 Gln Leu Gly Thr Tyr Asp Gly Thr Ile Tyr Asn Ala His Glu Val Val     290 295 300 Lys Arg Ile Gly His Leu Cys Asp Tyr Ile Glu Phe Asp Ser Ala Trp 305 310 315 320 Val Gly Tyr Glu Gln Phe Ile Pro Met Met Arg Asn Ser Ser Pro Leu                 325 330 335 Leu Ile Asp Asp Leu Gly Pro Glu Asp Pro Gly Ile Ile Val Val Gln             340 345 350 Ser Val His Lys Gln Gln Ala Gly Phe Ser Gln Thr Ser Gln Ile His         355 360 365 Lys Lys Asp Ser His Ile Lys Gly Gln Leu Arg Tyr Cys Asp His Lys     370 375 380 His Phe Asn Asn Ser Phe Asn Leu Phe Met Ser Thr Ser Pro Phe Tyr 385 390 395 400 Pro Met Tyr Ala Ala Leu Asp Val Asn Ala Ala Met Gln Glu Gly Glu                 405 410 415 Ala Gly Arg Lys Leu Trp His Asp Leu Leu Ile Thr Thr Ile Glu Ala             420 425 430 Arg Lys Lys Leu Ile Lys Ala Gly Ser Met Phe Arg Pro Phe Val Pro         435 440 445 Pro Val Val Asn Gly Lys Lys Trp Glu Asp Gly Asp Thr Glu Asp Met     450 455 460 Ala Asn Asn Ile Asp Tyr Trp Arg Phe Glu Lys Gly Ala Lys Trp His 465 470 475 480 Ala Tyr Glu Gly Tyr Gly Asp Asn Gln Tyr Tyr Val Asp Pro Asn Lys                 485 490 495 Phe Met Leu Thr Thr Pro Gly Ile Asn Pro Glu Thr Gly Asp Tyr Glu             500 505 510 Asp Phe Gly Val Pro Ala Thr Ile Val Ala Asn Tyr Leu Arg Asp His         515 520 525 Gly Ile Ile Pro Glu Lys Ser Asp Leu Asn Ser Ile Leu Phe Leu Met     530 535 540 Thr Pro Ala Glu Thr Pro Ala Lys Met Asn Asn Leu Ile Thr Gln Leu 545 550 555 560 Leu Gln Leu Gln Arg Leu Ile Glu Glu Asp Ala Pro Leu Lys Gln Val                 565 570 575 Leu Pro Ser Ile Tyr Ala Ala Asn Glu Glu Arg Tyr Asn Gly Tyr Thr             580 585 590 Ile Arg Glu Leu Cys Gln Glu Leu His Asp Phe Tyr Lys Asn Asn Asn         595 600 605 Thr Phe Thr Tyr Gln Lys Arg Leu Phe Leu Arg Glu Phe Phe Pro Glu     610 615 620 Gln Gly Met Leu Pro Tyr Glu Ala Arg Gln Glu Phe Ile Arg Asn His 625 630 635 640 Asn Lys Leu Val Pro Leu Asn Lys Ile Glu Gly Glu Ile Ala Leu Glu                 645 650 655 Gly Ala Leu Pro Tyr Pro Pro Gly Val Phe Cys Val Ala Pro Gly Glu             660 665 670 Lys Trp Ser Glu Thr Ala Val Lys Tyr Phe Thr Ile Leu Gln Asp Gly         675 680 685 Ile Asn Asn Phe Pro Gly Phe Glu Pro Glu Ile Gln Gly Val Tyr Phe     690 695 700 Lys Gln Glu Gly Asp Lys Val Val Leu Tyr Gly Glu Val Tyr Asp Ala 705 710 715 720 Glu Val Ala Lys Asn Asp Asp Arg Tyr Asn Asn                 725 730 <210> 6 <211> 731 <212> PRT <213> Artificial Sequence <220> <223> ORNITHINE DECARBOXYLASE MUTANT <400> 6 Met Ser Ser Ser Leu Lys Ile Ala Ser Thr Gln Glu Ala Arg Gln Tyr   1 5 10 15 Phe Asp Thr Asp Arg Val Val Val Asp Ala Val Gly Ser Asp Phe Thr              20 25 30 Asp Val Gly Ala Val Ile Ala Met Asp Tyr Glu Thr Asp Val Ile Asp          35 40 45 Ala Ala Asp Ala Thr Lys Phe Gly Ile Pro Val Phe Ala Val Thr Lys      50 55 60 Asp Ala Gln Ala Ile Ser Ala Asp Glu Leu Lys Lys Ile Phe His Ile  65 70 75 80 Ile Asp Leu Glu Asn Lys Phe Asp Ala Thr Val Asn Ala Arg Glu Ile                  85 90 95 Glu Thr Ala Val Asn Asn Tyr Glu Asp Ser Ile Leu Pro Pro Phe Phe             100 105 110 Lys Ser Leu Lys Glu Tyr Val Ser Arg Gly Leu Ile Gln Phe Asp Cys         115 120 125 Pro Gly His Gln Gly Gly Gln Tyr Tyr Arg Lys His Pro Ala Gly Arg     130 135 140 Glu Phe Tyr Asp Phe Phe Gly Glu Thr Val Phe Arg Ala Asp Leu Cys 145 150 155 160 Asn Ala Asp Val Ala Leu Gly Asp Leu Leu Ile His Glu Gly Pro Ala                 165 170 175 Val Ala Ala Glu Lys His Ala Ala Arg Val Tyr Asn Ala Asp Lys Thr             180 185 190 Tyr Phe Val Leu Gly Gly Ser Ser Asn Ala Asn Asn Thr Val Thr Ser         195 200 205 Ala Leu Val Ser Asn Gly Asp Leu Val Leu Phe Asp Arg Asn Asn His     210 215 220 Lys Ser Val Tyr Asn Ser Ala Leu Ala Met Ala Gly Gly Arg Pro Val 225 230 235 240 Tyr Leu Gln Thr Asn Arg Asn Pro Tyr Gly Phe Ile Gly Gly Ile Tyr                 245 250 255 Asp Ser Asp Phe Asp Glu Lys Lys Ile Arg Glu Leu Ala Ala Lys Val             260 265 270 Asp Pro Glu Arg Ala Lys Trp Lys Arg Pro Phe Arg Leu Ala Val Ile         275 280 285 Gln Leu Gly Thr Tyr Asp Gly Thr Ile Tyr Asn Ala His Glu Val Val     290 295 300 Lys Arg Ile Gly His Leu Cys Asp Tyr Ile Glu Phe Asp Ser Ala Trp 305 310 315 320 Val Gly Tyr Glu Gln Phe Ile Pro Met Met Arg Asn Ser Ser Pro Leu                 325 330 335 Leu Ile Asp Asp Leu Gly Pro Glu Asp Pro Gly Ile Ile Val Val Gln             340 345 350 Ser Val His Lys Gln Gln Ala Gly Phe Ser Gln Thr Ser Gln Ile His         355 360 365 Lys Lys Asp Ser His Ile Lys Gly Gln Leu Arg Tyr Cys Asp His Lys     370 375 380 His Phe Asn Asn Ser Phe Asn Leu Phe Met Ser Thr Ser Pro Phe Tyr 385 390 395 400 Pro Met Tyr Ala Ala Leu Asp Val Asn Ala Ala Met Gln Glu Gly Glu                 405 410 415 Ala Gly Arg Lys Leu Trp His Asp Leu Leu Ile Thr Thr Ile Glu Ala             420 425 430 Arg Lys Lys Leu Ile Lys Ala Gly Ser Met Phe Arg Pro Phe Val Pro         435 440 445 Pro Val Val Asn Gly Lys Lys Trp Glu Asp Gly Asp Thr Glu Asp Met     450 455 460 Ala Asn Asn Ile Asp Tyr Trp Arg Phe Glu Lys Gly Ala Lys Trp His 465 470 475 480 Ala Tyr Glu Gly Tyr Gly Asp Asn Gln Tyr Tyr Val Asp Pro Asn Lys                 485 490 495 Phe Met Leu Thr Thr Pro Gly Ile Asn Pro Glu Thr Gly Asp Tyr Glu             500 505 510 Asp Phe Gly Val Pro Ala Thr Ile Val Ala Asn Tyr Leu Arg Asp His         515 520 525 Gly Ile Ile Pro Glu Lys Ser Asp Leu Asn Ser Ile Leu Phe Leu Met     530 535 540 Thr Pro Ala Glu Thr Pro Ala Lys Met Asn Asn Leu Ile Thr Gln Leu 545 550 555 560 Leu Gln Leu Gln Arg Leu Ile Glu Glu Asp Ala Pro Leu Lys Gln Val                 565 570 575 Leu Pro Ser Ile Tyr Ala Ala Asn Glu Glu Arg Tyr Asn Gly Tyr Thr             580 585 590 Ile Arg Glu Leu Cys Gln Glu Leu His Asp Phe Tyr Lys Asn Asn Asn         595 600 605 Thr Phe Thr Tyr Gln Lys Arg Leu Phe Leu Arg Glu Phe Phe Pro Glu     610 615 620 Gln Gly Met Leu Pro Tyr Glu Ala Arg Gln Glu Phe Ile Arg Asn His 625 630 635 640 Asn Lys Leu Val Pro Leu Asn Lys Ile Glu Gly Glu Ile Ala Leu Glu                 645 650 655 Gly Ala Leu Pro Tyr Pro Pro Gly Val Phe Cys Val Ala Pro Gly Glu             660 665 670 Lys Trp Ser Glu Thr Ala Val Lys Tyr Phe Thr Ile Leu Gln Asp Gly         675 680 685 Ile Asn Asn Phe Pro Gly Phe Ala Pro Glu Ile Gln Gly Gly Tyr Phe     690 695 700 Lys Gln Glu Gly Asp Lys Val Val Leu Tyr Gly Glu Val Tyr Asp Ala 705 710 715 720 Glu Val Ala Lys Asn Asp Asp Arg Tyr Asn Asn                 725 730 <210> 7 <211> 731 <212> PRT <213> Artificial Sequence <220> <223> ORNITHINE DECARBOXYLASE MUTANT <400> 7 Met Ser Ser Ser Leu Lys Ile Ala Ser Thr Gln Glu Ala Arg Gln Tyr   1 5 10 15 Phe Asp Thr Asp Arg Val Val Val Asp Ala Val Gly Ser Asp Phe Thr              20 25 30 Asp Val Gly Ala Val Ile Ala Met Asp Tyr Glu Thr Asp Val Ile Asp          35 40 45 Ala Ala Asp Ala Thr Lys Phe Gly Ile Pro Val Phe Ala Val Thr Lys      50 55 60 Asp Ala Gln Ala Ile Ser Ala Asp Glu Leu Lys Lys Ile Phe His Ile  65 70 75 80 Ile Asp Leu Glu Asn Lys Phe Asp Ala Thr Val Asn Ala Arg Glu Ile                  85 90 95 Glu Thr Ala Val Asn Asn Tyr Glu Asp Ser Ile Leu Pro Pro Phe Phe             100 105 110 Lys Ser Leu Lys Glu Tyr Val Ser Arg Gly Leu Ile Gln Phe Asp Cys         115 120 125 Pro Gly His Gln Gly Gly Gln Tyr Tyr Arg Lys His Pro Ala Gly Arg     130 135 140 Glu Phe Tyr Asp Phe Phe Gly Glu Thr Val Phe Arg Ala Asp Leu Cys 145 150 155 160 Asn Ala Asp Val Ala Leu Gly Asp Leu Leu Ile His Glu Gly Pro Ala                 165 170 175 Val Ala Ala Glu Lys His Ala Ala Arg Val Tyr Asn Ala Asp Lys Thr             180 185 190 Tyr Phe Val Leu Gly Gly Ser Ser Asn Ala Asn Asn Thr Val Thr Ser         195 200 205 Ala Leu Val Ser Asn Gly Asp Leu Val Leu Phe Asp Arg Asn Asn His     210 215 220 Lys Ser Val Tyr Asn Ser Ala Leu Ala Met Ala Gly Gly Arg Pro Val 225 230 235 240 Tyr Leu Gln Thr Asn Arg Asn Pro Tyr Gly Phe Ile Gly Gly Ile Tyr                 245 250 255 Asp Ser Asp Phe Asp Glu Lys Lys Ile Arg Glu Leu Ala Ala Lys Val             260 265 270 Asp Pro Glu Arg Ala Lys Trp Lys Arg Pro Phe Arg Leu Ala Val Ile         275 280 285 Gln Leu Gly Thr Tyr Asp Gly Thr Ile Tyr Asn Ala His Glu Val Val     290 295 300 Lys Arg Ile Gly His Leu Cys Asp Tyr Ile Glu Phe Asp Ser Ala Trp 305 310 315 320 Val Gly Tyr Glu Gln Phe Ile Pro Met Met Arg Asn Ser Ser Pro Leu                 325 330 335 Leu Ile Asp Asp Leu Gly Pro Glu Asp Pro Gly Ile Ile Val Val Gln             340 345 350 Ser Val His Lys Gln Gln Ala Gly Phe Ser Gln Thr Ser Gln Ile His         355 360 365 Lys Lys Asp Ser His Ile Lys Gly Gln Leu Arg Tyr Cys Asp His Lys     370 375 380 His Phe Asn Asn Ser Phe Asn Leu Phe Met Ser Thr Ser Pro Phe Tyr 385 390 395 400 Pro Met Tyr Ala Ala Leu Asp Val Asn Ala Ala Met Gln Glu Gly Glu                 405 410 415 Ala Gly Arg Lys Leu Trp His Asp Leu Leu Ile Thr Thr Ile Glu Ala             420 425 430 Arg Lys Lys Leu Ile Lys Ala Gly Ser Met Phe Arg Pro Phe Val Pro         435 440 445 Pro Val Val Asn Gly Lys Lys Trp Glu Asp Gly Asp Thr Glu Asp Met     450 455 460 Ala Asn Asn Ile Asp Tyr Trp Arg Phe Glu Lys Gly Ala Lys Trp His 465 470 475 480 Ala Tyr Glu Gly Tyr Gly Asp Asn Gln Tyr Tyr Val Asp Pro Asn Lys                 485 490 495 Phe Met Leu Thr Thr Pro Gly Ile Asn Pro Glu Thr Gly Asp Tyr Glu             500 505 510 Asp Phe Gly Val Pro Ala Thr Ile Val Ala Asn Tyr Leu Arg Asp His         515 520 525 Gly Ile Ile Pro Glu Lys Ser Asp Leu Asn Ser Ile Leu Phe Leu Met     530 535 540 Thr Pro Ala Glu Thr Pro Ala Lys Met Asn Asn Leu Ile Thr Gln Leu 545 550 555 560 Leu Gln Leu Gln Arg Leu Ile Glu Glu Asp Ala Pro Leu Lys Gln Val                 565 570 575 Leu Pro Ser Ile Tyr Ala Ala Asn Glu Glu Arg Tyr Asn Gly Tyr Thr             580 585 590 Ile Arg Glu Leu Cys Gln Glu Leu His Asp Phe Tyr Lys Asn Asn Asn         595 600 605 Thr Phe Thr Tyr Gln Lys Arg Leu Phe Leu Arg Glu Phe Phe Pro Glu     610 615 620 Gln Gly Met Leu Pro Tyr Glu Ala Arg Gln Glu Phe Ile Arg Asn His 625 630 635 640 Asn Lys Leu Val Pro Leu Asn Lys Ile Glu Gly Glu Ile Ala Leu Glu                 645 650 655 Gly Ala Leu Pro Tyr Pro Pro Gly Val Phe Cys Val Ala Pro Gly Glu             660 665 670 Lys Trp Ser Glu Thr Ala Val Lys Tyr Phe Thr Ile Leu Gln Asp Gly         675 680 685 Ile Asn Asn Phe Pro Gly Phe Glu Pro Glu Ile Gln Gly Gly Tyr Phe     690 695 700 Lys Gln Glu Gly Asp Lys Val Val Leu Tyr Gly Glu Val Tyr Asp Ala 705 710 715 720 Glu Val Ala Lys Asn Asp Asp Arg Tyr Asn Asn                 725 730 <210> 8 <211> 731 <212> PRT <213> Artificial Sequence <220> <223> ORNITHINE DECARBOXYLASE MUTANT <400> 8 Met Ser Ser Ser Leu Lys Ile Ala Ser Thr Gln Glu Ala Arg Gln Tyr   1 5 10 15 Phe Asp Thr Asp Arg Val Val Val Asp Ala Val Gly Ser Asp Phe Thr              20 25 30 Asp Val Gly Ala Val Ile Ala Met Asp Tyr Glu Thr Asp Val Ile Asp          35 40 45 Ala Ala Asp Ala Thr Lys Phe Gly Ile Pro Val Phe Ala Val Thr Lys      50 55 60 Asp Ala Gln Ala Ile Ser Ala Asp Glu Leu Lys Lys Ile Phe His Ile  65 70 75 80 Ile Asp Leu Glu Asn Lys Phe Asp Ala Thr Val Asn Ala Arg Glu Ile                  85 90 95 Glu Thr Ala Val Asn Asn Tyr Glu Asp Ser Ile Leu Pro Pro Phe Phe             100 105 110 Lys Ser Leu Lys Glu Tyr Val Ser Arg Gly Leu Ile Gln Phe Asp Cys         115 120 125 Pro Gly His Gln Gly Gly Gln Tyr Tyr Arg Lys His Pro Ala Gly Arg     130 135 140 Glu Phe Tyr Asp Phe Phe Gly Glu Thr Val Phe Arg Ala Asp Leu Cys 145 150 155 160 Asn Ala Asp Val Ala Leu Gly Asp Leu Leu Ile His Glu Gly Pro Ala                 165 170 175 Val Ala Ala Glu Lys His Ala Ala Arg Val Tyr Asn Ala Asp Lys Thr             180 185 190 Tyr Phe Val Leu Gly Gly Ser Ser Asn Ala Asn Asn Thr Val Thr Ser         195 200 205 Ala Leu Val Ser Asn Gly Asp Leu Val Leu Phe Asp Arg Asn Asn His     210 215 220 Lys Ser Val Tyr Asn Ser Ala Leu Ala Met Ala Gly Gly Arg Pro Val 225 230 235 240 Tyr Leu Gln Thr Asn Arg Asn Pro Tyr Gly Phe Ile Gly Gly Ile Tyr                 245 250 255 Asp Ser Asp Phe Asp Glu Lys Lys Ile Arg Glu Leu Ala Ala Lys Val             260 265 270 Asp Pro Glu Arg Ala Lys Trp Lys Arg Pro Phe Arg Leu Ala Val Ile         275 280 285 Gln Leu Gly Thr Tyr Asp Gly Thr Ile Tyr Asn Ala His Glu Val Val     290 295 300 Lys Arg Ile Gly His Leu Cys Asp Tyr Ile Glu Phe Asp Ser Ala Trp 305 310 315 320 Val Gly Tyr Glu Gln Phe Ile Pro Met Met Arg Asn Ser Ser Pro Leu                 325 330 335 Leu Ile Asp Asp Leu Gly Pro Glu Asp Pro Gly Ile Ile Val Val Gln             340 345 350 Ser Val His Lys Gln Gln Ala Gly Phe Ser Gln Thr Ser Gln Ile His         355 360 365 Lys Lys Asp Ser His Ile Lys Gly Gln Leu Arg Tyr Cys Asp His Lys     370 375 380 His Phe Asn Asn Ser Phe Asn Leu Phe Met Ser Thr Ser Pro Phe Tyr 385 390 395 400 Pro Met Tyr Ala Ala Leu Asp Val Asn Ala Ala Met Gln Glu Gly Glu                 405 410 415 Ala Gly Arg Lys Leu Trp His Asp Leu Leu Ile Thr Thr Ile Glu Ala             420 425 430 Arg Lys Lys Leu Ile Lys Ala Gly Ser Met Phe Arg Pro Phe Val Pro         435 440 445 Pro Val Val Asn Gly Lys Lys Trp Glu Asp Gly Asp Thr Glu Asp Met     450 455 460 Ala Asn Asn Ile Asp Tyr Trp Arg Phe Glu Lys Gly Ala Lys Trp His 465 470 475 480 Ala Tyr Glu Gly Tyr Gly Asp Asn Gln Tyr Tyr Val Asp Pro Asn Lys                 485 490 495 Phe Met Leu Thr Thr Pro Gly Ile Asn Pro Glu Thr Gly Asp Tyr Glu             500 505 510 Asp Phe Gly Val Pro Ala Thr Ile Val Ala Asn Tyr Leu Arg Asp His         515 520 525 Gly Ile Ile Pro Glu Lys Ser Asp Leu Asn Ser Ile Leu Phe Leu Met     530 535 540 Thr Pro Ala Glu Thr Pro Ala Lys Met Asn Asn Leu Ile Thr Gln Leu 545 550 555 560 Leu Gln Leu Gln Arg Leu Ile Glu Glu Asp Ala Pro Leu Lys Gln Val                 565 570 575 Leu Pro Ser Ile Tyr Ala Ala Asn Glu Glu Arg Tyr Asn Gly Tyr Thr             580 585 590 Ile Arg Glu Leu Cys Gln Glu Leu His Asp Phe Tyr Lys Asn Asn Asn         595 600 605 Thr Phe Thr Tyr Gln Lys Arg Leu Phe Leu Arg Glu Phe Phe Pro Glu     610 615 620 Gln Gly Met Leu Pro Tyr Glu Ala Arg Gln Glu Phe Ile Arg Asn His 625 630 635 640 Asn Lys Leu Val Pro Leu Asn Lys Ile Glu Gly Glu Ile Ala Leu Glu                 645 650 655 Gly Ala Leu Pro Tyr Pro Pro Gly Val Phe Cys Val Ala Pro Gly Glu             660 665 670 Lys Trp Ser Glu Thr Ala Val Lys Tyr Phe Thr Ile Leu Gln Asp Gly         675 680 685 Ile Asn Asn Phe Pro Gly Phe Ala Pro Asp Ile Gln Gly Val Tyr Phe     690 695 700 Lys Gln Glu Gly Asp Lys Val Val Ala Tyr Gly Glu Val Tyr Asp Ala 705 710 715 720 Glu Val Ala Lys Asn Asp Asp Arg Tyr Asn Asn                 725 730 <210> 9 <211> 731 <212> PRT <213> Artificial Sequence <220> <223> ORNITHINE DECARBOXYLASE MUTANT <400> 9 Met Ser Ser Ser Leu Lys Ile Ala Ser Thr Gln Glu Ala Arg Gln Tyr   1 5 10 15 Phe Asp Thr Asp Arg Val Val Val Asp Ala Val Gly Ser Asp Phe Thr              20 25 30 Asp Val Gly Ala Val Ile Ala Met Asp Tyr Glu Thr Asp Val Ile Asp          35 40 45 Ala Ala Asp Ala Thr Lys Phe Gly Ile Pro Val Phe Ala Val Thr Lys      50 55 60 Asp Ala Gln Ala Ile Ser Ala Asp Glu Leu Lys Lys Ile Phe His Ile  65 70 75 80 Ile Asp Leu Glu Asn Lys Phe Asp Ala Thr Val Asn Ala Arg Glu Ile                  85 90 95 Glu Thr Ala Val Asn Asn Tyr Glu Asp Ser Ile Leu Pro Pro Phe Phe             100 105 110 Lys Ser Leu Lys Glu Tyr Val Ser Arg Gly Leu Ile Gln Phe Asp Cys         115 120 125 Pro Gly His Gln Gly Gly Gln Tyr Tyr Arg Lys His Pro Ala Gly Arg     130 135 140 Glu Phe Tyr Asp Phe Phe Gly Glu Thr Val Phe Arg Ala Asp Leu Cys 145 150 155 160 Asn Ala Asp Val Ala Leu Gly Asp Leu Leu Ile His Glu Gly Pro Ala                 165 170 175 Val Ala Ala Glu Lys His Ala Ala Arg Val Tyr Asn Ala Asp Lys Thr             180 185 190 Tyr Phe Val Leu Gly Gly Ser Ser Asn Ala Asn Asn Thr Val Thr Ser         195 200 205 Ala Leu Val Ser Asn Gly Asp Leu Val Leu Phe Asp Arg Asn Asn His     210 215 220 Lys Ser Val Tyr Asn Ser Ala Leu Ala Met Ala Gly Gly Arg Pro Val 225 230 235 240 Tyr Leu Gln Thr Asn Arg Asn Pro Tyr Gly Phe Ile Gly Gly Ile Tyr                 245 250 255 Asp Ser Asp Phe Asp Glu Lys Lys Ile Arg Glu Leu Ala Ala Lys Val             260 265 270 Asp Pro Glu Arg Ala Lys Trp Lys Arg Pro Phe Arg Leu Ala Val Ile         275 280 285 Gln Leu Gly Thr Tyr Asp Gly Thr Ile Tyr Asn Ala His Glu Val Val     290 295 300 Lys Arg Ile Gly His Leu Cys Asp Tyr Ile Glu Phe Asp Ser Ala Trp 305 310 315 320 Val Gly Tyr Glu Gln Phe Ile Pro Met Met Arg Asn Ser Ser Pro Leu                 325 330 335 Leu Ile Asp Asp Leu Gly Pro Glu Asp Pro Gly Ile Ile Val Val Gln             340 345 350 Ser Val His Lys Gln Gln Ala Gly Phe Ser Gln Thr Ser Gln Ile His         355 360 365 Lys Lys Asp Ser His Ile Lys Gly Gln Leu Arg Tyr Cys Asp His Lys     370 375 380 His Phe Asn Asn Ser Phe Asn Leu Phe Met Ser Thr Ser Pro Phe Tyr 385 390 395 400 Pro Met Tyr Ala Ala Leu Asp Val Asn Ala Ala Met Gln Glu Gly Glu                 405 410 415 Ala Gly Arg Lys Leu Trp His Asp Leu Leu Ile Thr Thr Ile Glu Ala             420 425 430 Arg Lys Lys Leu Ile Lys Ala Gly Ser Met Phe Arg Pro Phe Val Pro         435 440 445 Pro Val Val Asn Gly Lys Lys Trp Glu Asp Gly Asp Thr Glu Asp Met     450 455 460 Ala Asn Asn Ile Asp Tyr Trp Arg Phe Glu Lys Gly Ala Lys Trp His 465 470 475 480 Ala Tyr Glu Gly Tyr Gly Asp Asn Gln Tyr Tyr Val Asp Pro Asn Lys                 485 490 495 Phe Met Leu Thr Thr Pro Gly Ile Asn Pro Glu Thr Gly Asp Tyr Glu             500 505 510 Asp Phe Gly Val Pro Ala Thr Ile Val Ala Asn Tyr Leu Arg Asp His         515 520 525 Gly Ile Ile Pro Glu Lys Ser Asp Leu Asn Ser Ile Leu Phe Leu Met     530 535 540 Thr Pro Ala Glu Thr Pro Ala Lys Met Asn Asn Leu Ile Thr Gln Leu 545 550 555 560 Leu Gln Leu Gln Arg Leu Ile Glu Glu Asp Ala Pro Leu Lys Gln Val                 565 570 575 Leu Pro Ser Ile Tyr Ala Ala Asn Glu Glu Arg Tyr Asn Gly Tyr Thr             580 585 590 Ile Arg Glu Leu Cys Gln Glu Leu His Asp Phe Tyr Lys Asn Asn Asn         595 600 605 Thr Phe Thr Tyr Gln Lys Arg Leu Phe Leu Arg Glu Phe Phe Pro Glu     610 615 620 Gln Gly Met Leu Pro Tyr Glu Ala Arg Gln Glu Phe Ile Arg Asn His 625 630 635 640 Asn Lys Leu Val Pro Leu Asn Lys Ile Glu Gly Glu Ile Ala Leu Glu                 645 650 655 Gly Ala Leu Pro Tyr Pro Pro Gly Val Phe Cys Val Ala Pro Gly Glu             660 665 670 Lys Trp Ser Glu Thr Ala Val Lys Tyr Phe Thr Ile Leu Gln Asp Gly         675 680 685 Ile Asn Asn Phe Pro Gly Phe Ala Pro Asp Ile Gln Gly Val Tyr Phe     690 695 700 Lys Gln Glu Gly Asp Lys Val Val Leu Tyr Gly Glu Val Tyr Asp Ala 705 710 715 720 Glu Val Ala Lys Asn Asp Asp Arg Tyr Asn Asn                 725 730 <210> 10 <211> 2196 <212> DNA <213> Artificial Sequence <220> <223> A DNA encoding a amido acid sequences of SEQ ID NO.1 <400> 10 atgagttctt ctcttaaaat tgcttcgact caagaagcgc gtcaatattt cgatactgac 60 cgcgttgttg tcgatgctgt aggctctgat tttactgatg tcggtgctgt tatcgcaatg 120 gattacgaaa cagatgtcat cgacgctgct gatgcaacta agtttggtat tcctgttttt 180 gccgtaacta aggatgccca agctatcagt gctgatgagc tgaagaagat tttccacatc 240 attgatttgg aaaacaaatt tgatgctact gttaacgctc gtgaaatcga aactgctgtt 300 aacaactacg aagacagcat tttaccacca ttcttcaagt cattgaaaga atacgttagc 360 cgtggtttaa tccaattcga ctgcccaggt caccaaggtg gtcaatacta cagaaagcac 420 ccagctggtc gtgaattcta cgacttcttc ggcgaaactg tcttccgtgc agacttatgt 480 aacgctgacg ttgccttggg tgacttgctg atccacgaag gtcctgctgt tgctgctgaa 540 aagcatgctg cacgtgttta caacgctgac aagacttact tcgttttagg tggttcttcc 600 aacgctaaca acactgtaac atctgcttta gtttctaacg gcgacttggt attgttcgac 660 cggaacaacc acaagtccgt ttacaactca gctttagcta tggctggtgg ccgtcctgtt 720 tacctccaaa caaaccgtaa cccatacggc ttcatcggtg gtatctacga cagcgacttc 780 gatgaaaaga agatccgtga actggcagct aaggttgacc cagaacgtgc taagtggaag 840 cgtccattcc gtctggctgt tatccaatta ggtacttacg atggtactat ctacaacgca 900 cacgaagttg taaagcgtat cggtcacctt tgtgattaca tcgaattcga ctctgcttgg 960 gtaggttacg aacaattcat tcctatgatg cgtaactctt caccattatt gattgatgac 1020 cttggtccag aagatcctgg tatcattgtt gttcaatcag ttcacaagca acaagccggc 1080 ttctcacaaa cttcacaaat ccacaagaag gatagccaca tcaagggtca attacgttac 1140 tgtgaccaca agcactttaa caactccttc aacttgttca tgtctacttc accattctac 1200 ccaatgtatg cagcattaga cgttaacgct gctatgcaag aaggcgaagc aggtcgcaag 1260 ttatggcatg accttctgat tactaccatt gaagctcgta agaagttgat caaggctggc 1320 tcaatgttcc gtccattcgt tccacctgtt gttaacggca agaagtggga agatggcgac 1380 actgaagata tggctaacaa cattgactac tggcgctttg aaaagggtgc taagtggcat 1440 gcttacgaag gctacggcga caaccaatac tacgttgatc caaacaagtt catgttaact 1500 acacctggta tcaacccaga aactggtgac tacgaagact tcggtgttcc agctactatc 1560 gttgctaact acttacgtga ccacggtatc atccctgaaa agtctgactt gaactctatc 1620 ttgttcttga tgactccagc tgaaactcca gctaagatga acaacctgat cactcaactt 1680 cttcaattac aacgcttgat cgaagaagat gctccattga agcaagttct tccttcaatc 1740 tacgctgcta acgaagaacg ttacaatggc tacactatcc gtgaactttg ccaagaattg 1800 cacgacttct acaagaacaa caacacgttc acataccaga agcgtctctt cttacgtgaa 1860 ttcttcccag aacaaggtat gcttccatac gaagctcgtc aagaattcat ccgcaaccac 1920 aacaagcttg ttccattgaa caagatcgaa ggcgaaatcg ccctcgaagg tgctcttcca 1980 taccctccag gagtattctg tgtagcacca ggtgaaaagt ggtcagaaac tgctgttaag 2040 tacttcacta tcttacaaga tggtatcaac aacttccctg gattcgctcc agaaatccaa 2100 ggtgtatact tcaagcaaga aggcgacaag gttgttgctt acggtgaagt ttacgatgca 2160 gaagttgcta agaacgatga tcgttacaac aactaa 2196 <210> 11 <211> 2196 <212> DNA <213> Artificial Sequence <220> <223> A DNA encoding a amido acid sequences of SEQ ID NO.2 <400> 11 atgagttctt ctcttaaaat tgcttcgact caagaagcgc gtcaatattt cgatactgac 60 cgcgttgttg tcgatgctgt aggctctgat tttactgatg tcggtgctgt tatcgcaatg 120 gattacgaaa cagatgtcat cgacgctgct gatgcaacta agtttggtat tcctgttttt 180 gccgtaacta aggatgccca agctatcagt gctgatgagc tgaagaagat tttccacatc 240 attgatttgg aaaacaaatt tgatgctact gttaacgctc gtgaaatcga aactgctgtt 300 aacaactacg aagacagcat tttaccacca ttcttcaagt cattgaaaga atacgttagc 360 cgtggtttaa tccaattcga ctgcccaggt caccaaggtg gtcaatacta cagaaagcac 420 ccagctggtc gtgaattcta cgacttcttc ggcgaaactg tcttccgtgc agacttatgt 480 aacgctgacg ttgccttggg tgacttgctg atccacgaag gtcctgctgt tgctgctgaa 540 aagcatgctg cacgtgttta caacgctgac aagacttact tcgttttagg tggttcttcc 600 aacgctaaca acactgtaac atctgcttta gtttctaacg gcgacttggt attgttcgac 660 cggaacaacc acaagtccgt ttacaactca gctttagcta tggctggtgg ccgtcctgtt 720 tacctccaaa caaaccgtaa cccatacggc ttcatcggtg gtatctacga cagcgacttc 780 gatgaaaaga agatccgtga actggcagct aaggttgacc cagaacgtgc taagtggaag 840 cgtccattcc gtctggctgt tatccaatta ggtacttacg atggtactat ctacaacgca 900 cacgaagttg taaagcgtat cggtcacctt tgtgattaca tcgaattcga ctctgcttgg 960 gtaggttacg aacaattcat tcctatgatg cgtaactctt caccattatt gattgatgac 1020 cttggtccag aagatcctgg tatcattgtt gttcaatcag ttcacaagca acaagccggc 1080 ttctcacaaa cttcacaaat ccacaagaag gatagccaca tcaagggtca attacgttac 1140 tgtgaccaca agcactttaa caactccttc aacttgttca tgtctacttc accattctac 1200 ccaatgtatg cagcattaga cgttaacgct gctatgcaag aaggcgaagc aggtcgcaag 1260 ttatggcatg accttctgat tactaccatt gaagctcgta agaagttgat caaggctggc 1320 tcaatgttcc gtccattcgt tccacctgtt gttaacggca agaagtggga agatggcgac 1380 actgaagata tggctaacaa cattgactac tggcgctttg aaaagggtgc taagtggcat 1440 gcttacgaag gctacggcga caaccaatac tacgttgatc caaacaagtt catgttaact 1500 acacctggta tcaacccaga aactggtgac tacgaagact tcggtgttcc agctactatc 1560 gttgctaact acttacgtga ccacggtatc atccctgaaa agtctgactt gaactctatc 1620 ttgttcttga tgactccagc tgaaactcca gctaagatga acaacctgat cactcaactt 1680 cttcaattac aacgcttgat cgaagaagat gctccattga agcaagttct tccttcaatc 1740 tacgctgcta acgaagaacg ttacaatggc tacactatcc gtgaactttg ccaagaattg 1800 cacgacttct acaagaacaa caacacgttc acataccaga agcgtctctt cttacgtgaa 1860 ttcttcccag aacaaggtat gcttccatac gaagctcgtc aagaattcat ccgcaaccac 1920 aacaagcttg ttccattgaa caagatcgaa ggcgaaatcg ccctcgaagg tgctcttcca 1980 taccctccag gagtattctg tgtagcacca ggtgaaaagt ggtcagaaac tgctgttaag 2040 tacttcacta tcttacaaga tggtatcaac aacttccctg gattcgaacc agaaatccaa 2100 ggtgtatact tcaagcaaga aggcgacaag gttgttgctt acggtgaagt ttacgatgca 2160 gaagttgcta agaacgatga tcgttacaac aactaa 2196 <210> 12 <211> 2196 <212> DNA <213> Artificial Sequence <220> <223> A DNA encoding a amido acid sequences of SEQ ID NO.3 <400> 12 atgagttctt ctcttaaaat tgcttcgact caagaagcgc gtcaatattt cgatactgac 60 cgcgttgttg tcgatgctgt aggctctgat tttactgatg tcggtgctgt tatcgcaatg 120 gattacgaaa cagatgtcat cgacgctgct gatgcaacta agtttggtat tcctgttttt 180 gccgtaacta aggatgccca agctatcagt gctgatgagc tgaagaagat tttccacatc 240 attgatttgg aaaacaaatt tgatgctact gttaacgctc gtgaaatcga aactgctgtt 300 aacaactacg aagacagcat tttaccacca ttcttcaagt cattgaaaga atacgttagc 360 cgtggtttaa tccaattcga ctgcccaggt caccaaggtg gtcaatacta cagaaagcac 420 ccagctggtc gtgaattcta cgacttcttc ggcgaaactg tcttccgtgc agacttatgt 480 aacgctgacg ttgccttggg tgacttgctg atccacgaag gtcctgctgt tgctgctgaa 540 aagcatgctg cacgtgttta caacgctgac aagacttact tcgttttagg tggttcttcc 600 aacgctaaca acactgtaac atctgcttta gtttctaacg gcgacttggt attgttcgac 660 cggaacaacc acaagtccgt ttacaactca gctttagcta tggctggtgg ccgtcctgtt 720 tacctccaaa caaaccgtaa cccatacggc ttcatcggtg gtatctacga cagcgacttc 780 gatgaaaaga agatccgtga actggcagct aaggttgacc cagaacgtgc taagtggaag 840 cgtccattcc gtctggctgt tatccaatta ggtacttacg atggtactat ctacaacgca 900 cacgaagttg taaagcgtat cggtcacctt tgtgattaca tcgaattcga ctctgcttgg 960 gtaggttacg aacaattcat tcctatgatg cgtaactctt caccattatt gattgatgac 1020 cttggtccag aagatcctgg tatcattgtt gttcaatcag ttcacaagca acaagccggc 1080 ttctcacaaa cttcacaaat ccacaagaag gatagccaca tcaagggtca attacgttac 1140 tgtgaccaca agcactttaa caactccttc aacttgttca tgtctacttc accattctac 1200 ccaatgtatg cagcattaga cgttaacgct gctatgcaag aaggcgaagc aggtcgcaag 1260 ttatggcatg accttctgat tactaccatt gaagctcgta agaagttgat caaggctggc 1320 tcaatgttcc gtccattcgt tccacctgtt gttaacggca agaagtggga agatggcgac 1380 actgaagata tggctaacaa cattgactac tggcgctttg aaaagggtgc taagtggcat 1440 gcttacgaag gctacggcga caaccaatac tacgttgatc caaacaagtt catgttaact 1500 acacctggta tcaacccaga aactggtgac tacgaagact tcggtgttcc agctactatc 1560 gttgctaact acttacgtga ccacggtatc atccctgaaa agtctgactt gaactctatc 1620 ttgttcttga tgactccagc tgaaactcca gctaagatga acaacctgat cactcaactt 1680 cttcaattac aacgcttgat cgaagaagat gctccattga agcaagttct tccttcaatc 1740 tacgctgcta acgaagaacg ttacaatggc tacactatcc gtgaactttg ccaagaattg 1800 cacgacttct acaagaacaa caacacgttc acataccaga agcgtctctt cttacgtgaa 1860 ttcttcccag aacaaggtat gcttccatac gaagctcgtc aagaattcat ccgcaaccac 1920 aacaagcttg ttccattgaa caagatcgaa ggcgaaatcg ccctcgaagg tgctcttcca 1980 taccctccag gagtattctg tgtagcacca ggtgaaaagt ggtcagaaac tgctgttaag 2040 tacttcacta tcttacaaga tggtatcaac aacttccctg gattcgctcc agaaatccaa 2100 ggtgggtact tcaagcaaga aggcgacaag gttgttgctt acggtgaagt ttacgatgca 2160 gaagttgcta agaacgatga tcgttacaac aactaa 2196 <210> 13 <211> 2196 <212> DNA <213> Artificial Sequence <220> <223> A DNA encoding a amido acid sequences of SEQ ID NO.4 <400> 13 atgagttctt ctcttaaaat tgcttcgact caagaagcgc gtcaatattt cgatactgac 60 cgcgttgttg tcgatgctgt aggctctgat tttactgatg tcggtgctgt tatcgcaatg 120 gattacgaaa cagatgtcat cgacgctgct gatgcaacta agtttggtat tcctgttttt 180 gccgtaacta aggatgccca agctatcagt gctgatgagc tgaagaagat tttccacatc 240 attgatttgg aaaacaaatt tgatgctact gttaacgctc gtgaaatcga aactgctgtt 300 aacaactacg aagacagcat tttaccacca ttcttcaagt cattgaaaga atacgttagc 360 cgtggtttaa tccaattcga ctgcccaggt caccaaggtg gtcaatacta cagaaagcac 420 ccagctggtc gtgaattcta cgacttcttc ggcgaaactg tcttccgtgc agacttatgt 480 aacgctgacg ttgccttggg tgacttgctg atccacgaag gtcctgctgt tgctgctgaa 540 aagcatgctg cacgtgttta caacgctgac aagacttact tcgttttagg tggttcttcc 600 aacgctaaca acactgtaac atctgcttta gtttctaacg gcgacttggt attgttcgac 660 cggaacaacc acaagtccgt ttacaactca gctttagcta tggctggtgg ccgtcctgtt 720 tacctccaaa caaaccgtaa cccatacggc ttcatcggtg gtatctacga cagcgacttc 780 gatgaaaaga agatccgtga actggcagct aaggttgacc cagaacgtgc taagtggaag 840 cgtccattcc gtctggctgt tatccaatta ggtacttacg atggtactat ctacaacgca 900 cacgaagttg taaagcgtat cggtcacctt tgtgattaca tcgaattcga ctctgcttgg 960 gtaggttacg aacaattcat tcctatgatg cgtaactctt caccattatt gattgatgac 1020 cttggtccag aagatcctgg tatcattgtt gttcaatcag ttcacaagca acaagccggc 1080 ttctcacaaa cttcacaaat ccacaagaag gatagccaca tcaagggtca attacgttac 1140 tgtgaccaca agcactttaa caactccttc aacttgttca tgtctacttc accattctac 1200 ccaatgtatg cagcattaga cgttaacgct gctatgcaag aaggcgaagc aggtcgcaag 1260 ttatggcatg accttctgat tactaccatt gaagctcgta agaagttgat caaggctggc 1320 tcaatgttcc gtccattcgt tccacctgtt gttaacggca agaagtggga agatggcgac 1380 actgaagata tggctaacaa cattgactac tggcgctttg aaaagggtgc taagtggcat 1440 gcttacgaag gctacggcga caaccaatac tacgttgatc caaacaagtt catgttaact 1500 acacctggta tcaacccaga aactggtgac tacgaagact tcggtgttcc agctactatc 1560 gttgctaact acttacgtga ccacggtatc atccctgaaa agtctgactt gaactctatc 1620 ttgttcttga tgactccagc tgaaactcca gctaagatga acaacctgat cactcaactt 1680 cttcaattac aacgcttgat cgaagaagat gctccattga agcaagttct tccttcaatc 1740 tacgctgcta acgaagaacg ttacaatggc tacactatcc gtgaactttg ccaagaattg 1800 cacgacttct acaagaacaa caacacgttc acataccaga agcgtctctt cttacgtgaa 1860 ttcttcccag aacaaggtat gcttccatac gaagctcgtc aagaattcat ccgcaaccac 1920 aacaagcttg ttccattgaa caagatcgaa ggcgaaatcg ccctcgaagg tgctcttcca 1980 taccctccag gagtattctg tgtagcacca ggtgaaaagt ggtcagaaac tgctgttaag 2040 tacttcacta tcttacaaga tggtatcaac aacttccctg gattcgctcc agaaatccaa 2100 ggtgtatact tcaagcaaga aggcgacaag gttgttcttt acggtgaagt ttacgatgca 2160 gaagttgcta agaacgatga tcgttacaac aactaa 2196 <210> 14 <211> 2196 <212> DNA <213> Artificial Sequence <220> <223> A DNA encoding a amido acid sequences of SEQ ID NO.5 <400> 14 atgagttctt ctcttaaaat tgcttcgact caagaagcgc gtcaatattt cgatactgac 60 cgcgttgttg tcgatgctgt aggctctgat tttactgatg tcggtgctgt tatcgcaatg 120 gattacgaaa cagatgtcat cgacgctgct gatgcaacta agtttggtat tcctgttttt 180 gccgtaacta aggatgccca agctatcagt gctgatgagc tgaagaagat tttccacatc 240 attgatttgg aaaacaaatt tgatgctact gttaacgctc gtgaaatcga aactgctgtt 300 aacaactacg aagacagcat tttaccacca ttcttcaagt cattgaaaga atacgttagc 360 cgtggtttaa tccaattcga ctgcccaggt caccaaggtg gtcaatacta cagaaagcac 420 ccagctggtc gtgaattcta cgacttcttc ggcgaaactg tcttccgtgc agacttatgt 480 aacgctgacg ttgccttggg tgacttgctg atccacgaag gtcctgctgt tgctgctgaa 540 aagcatgctg cacgtgttta caacgctgac aagacttact tcgttttagg tggttcttcc 600 aacgctaaca acactgtaac atctgcttta gtttctaacg gcgacttggt attgttcgac 660 cggaacaacc acaagtccgt ttacaactca gctttagcta tggctggtgg ccgtcctgtt 720 tacctccaaa caaaccgtaa cccatacggc ttcatcggtg gtatctacga cagcgacttc 780 gatgaaaaga agatccgtga actggcagct aaggttgacc cagaacgtgc taagtggaag 840 cgtccattcc gtctggctgt tatccaatta ggtacttacg atggtactat ctacaacgca 900 cacgaagttg taaagcgtat cggtcacctt tgtgattaca tcgaattcga ctctgcttgg 960 gtaggttacg aacaattcat tcctatgatg cgtaactctt caccattatt gattgatgac 1020 cttggtccag aagatcctgg tatcattgtt gttcaatcag ttcacaagca acaagccggc 1080 ttctcacaaa cttcacaaat ccacaagaag gatagccaca tcaagggtca attacgttac 1140 tgtgaccaca agcactttaa caactccttc aacttgttca tgtctacttc accattctac 1200 ccaatgtatg cagcattaga cgttaacgct gctatgcaag aaggcgaagc aggtcgcaag 1260 ttatggcatg accttctgat tactaccatt gaagctcgta agaagttgat caaggctggc 1320 tcaatgttcc gtccattcgt tccacctgtt gttaacggca agaagtggga agatggcgac 1380 actgaagata tggctaacaa cattgactac tggcgctttg aaaagggtgc taagtggcat 1440 gcttacgaag gctacggcga caaccaatac tacgttgatc caaacaagtt catgttaact 1500 acacctggta tcaacccaga aactggtgac tacgaagact tcggtgttcc agctactatc 1560 gttgctaact acttacgtga ccacggtatc atccctgaaa agtctgactt gaactctatc 1620 ttgttcttga tgactccagc tgaaactcca gctaagatga acaacctgat cactcaactt 1680 cttcaattac aacgcttgat cgaagaagat gctccattga agcaagttct tccttcaatc 1740 tacgctgcta acgaagaacg ttacaatggc tacactatcc gtgaactttg ccaagaattg 1800 cacgacttct acaagaacaa caacacgttc acataccaga agcgtctctt cttacgtgaa 1860 ttcttcccag aacaaggtat gcttccatac gaagctcgtc aagaattcat ccgcaaccac 1920 aacaagcttg ttccattgaa caagatcgaa ggcgaaatcg ccctcgaagg tgctcttcca 1980 taccctccag gagtattctg tgtagcacca ggtgaaaagt ggtcagaaac tgctgttaag 2040 tacttcacta tcttacaaga tggtatcaac aacttccctg gattcgaacc agaaatccaa 2100 ggtgtatact tcaagcaaga aggcgacaag gttgttcttt acggtgaagt ttacgatgca 2160 gaagttgcta agaacgatga tcgttacaac aactaa 2196 <210> 15 <211> 2196 <212> DNA <213> Artificial Sequence <220> <223> A DNA encoding a amido acid sequences of SEQ ID NO.6 <400> 15 atgagttctt ctcttaaaat tgcttcgact caagaagcgc gtcaatattt cgatactgac 60 cgcgttgttg tcgatgctgt aggctctgat tttactgatg tcggtgctgt tatcgcaatg 120 gattacgaaa cagatgtcat cgacgctgct gatgcaacta agtttggtat tcctgttttt 180 gccgtaacta aggatgccca agctatcagt gctgatgagc tgaagaagat tttccacatc 240 attgatttgg aaaacaaatt tgatgctact gttaacgctc gtgaaatcga aactgctgtt 300 aacaactacg aagacagcat tttaccacca ttcttcaagt cattgaaaga atacgttagc 360 cgtggtttaa tccaattcga ctgcccaggt caccaaggtg gtcaatacta cagaaagcac 420 ccagctggtc gtgaattcta cgacttcttc ggcgaaactg tcttccgtgc agacttatgt 480 aacgctgacg ttgccttggg tgacttgctg atccacgaag gtcctgctgt tgctgctgaa 540 aagcatgctg cacgtgttta caacgctgac aagacttact tcgttttagg tggttcttcc 600 aacgctaaca acactgtaac atctgcttta gtttctaacg gcgacttggt attgttcgac 660 cggaacaacc acaagtccgt ttacaactca gctttagcta tggctggtgg ccgtcctgtt 720 tacctccaaa caaaccgtaa cccatacggc ttcatcggtg gtatctacga cagcgacttc 780 gatgaaaaga agatccgtga actggcagct aaggttgacc cagaacgtgc taagtggaag 840 cgtccattcc gtctggctgt tatccaatta ggtacttacg atggtactat ctacaacgca 900 cacgaagttg taaagcgtat cggtcacctt tgtgattaca tcgaattcga ctctgcttgg 960 gtaggttacg aacaattcat tcctatgatg cgtaactctt caccattatt gattgatgac 1020 cttggtccag aagatcctgg tatcattgtt gttcaatcag ttcacaagca acaagccggc 1080 ttctcacaaa cttcacaaat ccacaagaag gatagccaca tcaagggtca attacgttac 1140 tgtgaccaca agcactttaa caactccttc aacttgttca tgtctacttc accattctac 1200 ccaatgtatg cagcattaga cgttaacgct gctatgcaag aaggcgaagc aggtcgcaag 1260 ttatggcatg accttctgat tactaccatt gaagctcgta agaagttgat caaggctggc 1320 tcaatgttcc gtccattcgt tccacctgtt gttaacggca agaagtggga agatggcgac 1380 actgaagata tggctaacaa cattgactac tggcgctttg aaaagggtgc taagtggcat 1440 gcttacgaag gctacggcga caaccaatac tacgttgatc caaacaagtt catgttaact 1500 acacctggta tcaacccaga aactggtgac tacgaagact tcggtgttcc agctactatc 1560 gttgctaact acttacgtga ccacggtatc atccctgaaa agtctgactt gaactctatc 1620 ttgttcttga tgactccagc tgaaactcca gctaagatga acaacctgat cactcaactt 1680 cttcaattac aacgcttgat cgaagaagat gctccattga agcaagttct tccttcaatc 1740 tacgctgcta acgaagaacg ttacaatggc tacactatcc gtgaactttg ccaagaattg 1800 cacgacttct acaagaacaa caacacgttc acataccaga agcgtctctt cttacgtgaa 1860 ttcttcccag aacaaggtat gcttccatac gaagctcgtc aagaattcat ccgcaaccac 1920 aacaagcttg ttccattgaa caagatcgaa ggcgaaatcg ccctcgaagg tgctcttcca 1980 taccctccag gagtattctg tgtagcacca ggtgaaaagt ggtcagaaac tgctgttaag 2040 tacttcacta tcttacaaga tggtatcaac aacttccctg gattcgctcc agaaatccaa 2100 ggtgggtact tcaagcaaga aggcgacaag gttgttcttt acggtgaagt ttacgatgca 2160 gaagttgcta agaacgatga tcgttacaac aactaa 2196 <210> 16 <211> 2196 <212> DNA <213> Artificial Sequence <220> <223> A DNA encoding a amido acid sequences of SEQ ID NO.7 <400> 16 atgagttctt ctcttaaaat tgcttcgact caagaagcgc gtcaatattt cgatactgac 60 cgcgttgttg tcgatgctgt aggctctgat tttactgatg tcggtgctgt tatcgcaatg 120 gattacgaaa cagatgtcat cgacgctgct gatgcaacta agtttggtat tcctgttttt 180 gccgtaacta aggatgccca agctatcagt gctgatgagc tgaagaagat tttccacatc 240 attgatttgg aaaacaaatt tgatgctact gttaacgctc gtgaaatcga aactgctgtt 300 aacaactacg aagacagcat tttaccacca ttcttcaagt cattgaaaga atacgttagc 360 cgtggtttaa tccaattcga ctgcccaggt caccaaggtg gtcaatacta cagaaagcac 420 ccagctggtc gtgaattcta cgacttcttc ggcgaaactg tcttccgtgc agacttatgt 480 aacgctgacg ttgccttggg tgacttgctg atccacgaag gtcctgctgt tgctgctgaa 540 aagcatgctg cacgtgttta caacgctgac aagacttact tcgttttagg tggttcttcc 600 aacgctaaca acactgtaac atctgcttta gtttctaacg gcgacttggt attgttcgac 660 cggaacaacc acaagtccgt ttacaactca gctttagcta tggctggtgg ccgtcctgtt 720 tacctccaaa caaaccgtaa cccatacggc ttcatcggtg gtatctacga cagcgacttc 780 gatgaaaaga agatccgtga actggcagct aaggttgacc cagaacgtgc taagtggaag 840 cgtccattcc gtctggctgt tatccaatta ggtacttacg atggtactat ctacaacgca 900 cacgaagttg taaagcgtat cggtcacctt tgtgattaca tcgaattcga ctctgcttgg 960 gtaggttacg aacaattcat tcctatgatg cgtaactctt caccattatt gattgatgac 1020 cttggtccag aagatcctgg tatcattgtt gttcaatcag ttcacaagca acaagccggc 1080 ttctcacaaa cttcacaaat ccacaagaag gatagccaca tcaagggtca attacgttac 1140 tgtgaccaca agcactttaa caactccttc aacttgttca tgtctacttc accattctac 1200 ccaatgtatg cagcattaga cgttaacgct gctatgcaag aaggcgaagc aggtcgcaag 1260 ttatggcatg accttctgat tactaccatt gaagctcgta agaagttgat caaggctggc 1320 tcaatgttcc gtccattcgt tccacctgtt gttaacggca agaagtggga agatggcgac 1380 actgaagata tggctaacaa cattgactac tggcgctttg aaaagggtgc taagtggcat 1440 gcttacgaag gctacggcga caaccaatac tacgttgatc caaacaagtt catgttaact 1500 acacctggta tcaacccaga aactggtgac tacgaagact tcggtgttcc agctactatc 1560 gttgctaact acttacgtga ccacggtatc atccctgaaa agtctgactt gaactctatc 1620 ttgttcttga tgactccagc tgaaactcca gctaagatga acaacctgat cactcaactt 1680 cttcaattac aacgcttgat cgaagaagat gctccattga agcaagttct tccttcaatc 1740 tacgctgcta acgaagaacg ttacaatggc tacactatcc gtgaactttg ccaagaattg 1800 cacgacttct acaagaacaa caacacgttc acataccaga agcgtctctt cttacgtgaa 1860 ttcttcccag aacaaggtat gcttccatac gaagctcgtc aagaattcat ccgcaaccac 1920 aacaagcttg ttccattgaa caagatcgaa ggcgaaatcg ccctcgaagg tgctcttcca 1980 taccctccag gagtattctg tgtagcacca ggtgaaaagt ggtcagaaac tgctgttaag 2040 tacttcacta tcttacaaga tggtatcaac aacttccctg gattcgaacc agaaatccaa 2100 ggtgggtact tcaagcaaga aggcgacaag gttgttcttt acggtgaagt ttacgatgca 2160 gaagttgcta agaacgatga tcgttacaac aactaa 2196 <210> 17 <211> 2196 <212> DNA <213> Artificial Sequence <220> <223> A DNA encoding a amido acid sequences of SEQ ID NO.8 <400> 17 atgagttctt ctcttaaaat tgcttcgact caagaagcgc gtcaatattt cgatactgac 60 cgcgttgttg tcgatgctgt aggctctgat tttactgatg tcggtgctgt tatcgcaatg 120 gattacgaaa cagatgtcat cgacgctgct gatgcaacta agtttggtat tcctgttttt 180 gccgtaacta aggatgccca agctatcagt gctgatgagc tgaagaagat tttccacatc 240 attgatttgg aaaacaaatt tgatgctact gttaacgctc gtgaaatcga aactgctgtt 300 aacaactacg aagacagcat tttaccacca ttcttcaagt cattgaaaga atacgttagc 360 cgtggtttaa tccaattcga ctgcccaggt caccaaggtg gtcaatacta cagaaagcac 420 ccagctggtc gtgaattcta cgacttcttc ggcgaaactg tcttccgtgc agacttatgt 480 aacgctgacg ttgccttggg tgacttgctg atccacgaag gtcctgctgt tgctgctgaa 540 aagcatgctg cacgtgttta caacgctgac aagacttact tcgttttagg tggttcttcc 600 aacgctaaca acactgtaac atctgcttta gtttctaacg gcgacttggt attgttcgac 660 cggaacaacc acaagtccgt ttacaactca gctttagcta tggctggtgg ccgtcctgtt 720 tacctccaaa caaaccgtaa cccatacggc ttcatcggtg gtatctacga cagcgacttc 780 gatgaaaaga agatccgtga actggcagct aaggttgacc cagaacgtgc taagtggaag 840 cgtccattcc gtctggctgt tatccaatta ggtacttacg atggtactat ctacaacgca 900 cacgaagttg taaagcgtat cggtcacctt tgtgattaca tcgaattcga ctctgcttgg 960 gtaggttacg aacaattcat tcctatgatg cgtaactctt caccattatt gattgatgac 1020 cttggtccag aagatcctgg tatcattgtt gttcaatcag ttcacaagca acaagccggc 1080 ttctcacaaa cttcacaaat ccacaagaag gatagccaca tcaagggtca attacgttac 1140 tgtgaccaca agcactttaa caactccttc aacttgttca tgtctacttc accattctac 1200 ccaatgtatg cagcattaga cgttaacgct gctatgcaag aaggcgaagc aggtcgcaag 1260 ttatggcatg accttctgat tactaccatt gaagctcgta agaagttgat caaggctggc 1320 tcaatgttcc gtccattcgt tccacctgtt gttaacggca agaagtggga agatggcgac 1380 actgaagata tggctaacaa cattgactac tggcgctttg aaaagggtgc taagtggcat 1440 gcttacgaag gctacggcga caaccaatac tacgttgatc caaacaagtt catgttaact 1500 acacctggta tcaacccaga aactggtgac tacgaagact tcggtgttcc agctactatc 1560 gttgctaact acttacgtga ccacggtatc atccctgaaa agtctgactt gaactctatc 1620 ttgttcttga tgactccagc tgaaactcca gctaagatga acaacctgat cactcaactt 1680 cttcaattac aacgcttgat cgaagaagat gctccattga agcaagttct tccttcaatc 1740 tacgctgcta acgaagaacg ttacaatggc tacactatcc gtgaactttg ccaagaattg 1800 cacgacttct acaagaacaa caacacgttc acataccaga agcgtctctt cttacgtgaa 1860 ttcttcccag aacaaggtat gcttccatac gaagctcgtc aagaattcat ccgcaaccac 1920 aacaagcttg ttccattgaa caagatcgaa ggcgaaatcg ccctcgaagg tgctcttcca 1980 taccctccag gagtattctg tgtagcacca ggtgaaaagt ggtcagaaac tgctgttaag 2040 tacttcacta tcttacaaga tggtatcaac aacttccctg gattcgctcc agatatccaa 2100 ggtgtatact tcaagcaaga aggcgacaag gttgttgctt acggtgaagt ttacgatgca 2160 gaagttgcta agaacgatga tcgttacaac aactaa 2196 <210> 18 <211> 2196 <212> DNA <213> Artificial Sequence <220> <223> A DNA encoding a amido acid sequences of SEQ ID NO.9 <400> 18 atgagttctt ctcttaaaat tgcttcgact caagaagcgc gtcaatattt cgatactgac 60 cgcgttgttg tcgatgctgt aggctctgat tttactgatg tcggtgctgt tatcgcaatg 120 gattacgaaa cagatgtcat cgacgctgct gatgcaacta agtttggtat tcctgttttt 180 gccgtaacta aggatgccca agctatcagt gctgatgagc tgaagaagat tttccacatc 240 attgatttgg aaaacaaatt tgatgctact gttaacgctc gtgaaatcga aactgctgtt 300 aacaactacg aagacagcat tttaccacca ttcttcaagt cattgaaaga atacgttagc 360 cgtggtttaa tccaattcga ctgcccaggt caccaaggtg gtcaatacta cagaaagcac 420 ccagctggtc gtgaattcta cgacttcttc ggcgaaactg tcttccgtgc agacttatgt 480 aacgctgacg ttgccttggg tgacttgctg atccacgaag gtcctgctgt tgctgctgaa 540 aagcatgctg cacgtgttta caacgctgac aagacttact tcgttttagg tggttcttcc 600 aacgctaaca acactgtaac atctgcttta gtttctaacg gcgacttggt attgttcgac 660 cggaacaacc acaagtccgt ttacaactca gctttagcta tggctggtgg ccgtcctgtt 720 tacctccaaa caaaccgtaa cccatacggc ttcatcggtg gtatctacga cagcgacttc 780 gatgaaaaga agatccgtga actggcagct aaggttgacc cagaacgtgc taagtggaag 840 cgtccattcc gtctggctgt tatccaatta ggtacttacg atggtactat ctacaacgca 900 cacgaagttg taaagcgtat cggtcacctt tgtgattaca tcgaattcga ctctgcttgg 960 gtaggttacg aacaattcat tcctatgatg cgtaactctt caccattatt gattgatgac 1020 cttggtccag aagatcctgg tatcattgtt gttcaatcag ttcacaagca acaagccggc 1080 ttctcacaaa cttcacaaat ccacaagaag gatagccaca tcaagggtca attacgttac 1140 tgtgaccaca agcactttaa caactccttc aacttgttca tgtctacttc accattctac 1200 ccaatgtatg cagcattaga cgttaacgct gctatgcaag aaggcgaagc aggtcgcaag 1260 ttatggcatg accttctgat tactaccatt gaagctcgta agaagttgat caaggctggc 1320 tcaatgttcc gtccattcgt tccacctgtt gttaacggca agaagtggga agatggcgac 1380 actgaagata tggctaacaa cattgactac tggcgctttg aaaagggtgc taagtggcat 1440 gcttacgaag gctacggcga caaccaatac tacgttgatc caaacaagtt catgttaact 1500 acacctggta tcaacccaga aactggtgac tacgaagact tcggtgttcc agctactatc 1560 gttgctaact acttacgtga ccacggtatc atccctgaaa agtctgactt gaactctatc 1620 ttgttcttga tgactccagc tgaaactcca gctaagatga acaacctgat cactcaactt 1680 cttcaattac aacgcttgat cgaagaagat gctccattga agcaagttct tccttcaatc 1740 tacgctgcta acgaagaacg ttacaatggc tacactatcc gtgaactttg ccaagaattg 1800 cacgacttct acaagaacaa caacacgttc acataccaga agcgtctctt cttacgtgaa 1860 ttcttcccag aacaaggtat gcttccatac gaagctcgtc aagaattcat ccgcaaccac 1920 aacaagcttg ttccattgaa caagatcgaa ggcgaaatcg ccctcgaagg tgctcttcca 1980 taccctccag gagtattctg tgtagcacca ggtgaaaagt ggtcagaaac tgctgttaag 2040 tacttcacta tcttacaaga tggtatcaac aacttccctg gattcgctcc agatatccaa 2100 ggtgtatact tcaagcaaga aggcgacaag gttgttcttt acggtgaagt ttacgatgca 2160 gaagttgcta agaacgatga tcgttacaac aactaa 2196

Claims (9)

Variants of ornithine decarboxylase represented by any one of the following amino acid sequences:
In the amino acid sequence of SEQ ID 1, the amino acid sequence of the 713th alanine substituted for leucine; or
Amino acid sequence of 698th glutamic acid with aspartic acid and 713th alanine with leucine in amino acid sequence of SEQ ID NO: 1. delete DNA encoding a variant of ornithine decarboxylase according to claim 1. The method of claim 3, wherein
DNA is represented by SEQ ID NO: 13 or 18. A recombinant DNA vector comprising the DNA according to claim 3. A host cell transformed with the recombinant DNA vector according to claim 5. A method for producing putrescine, wherein any one selected from the group consisting of a host cell transformed with the recombinant DNA vector according to claim 6 and an extract of the host cell is used as a biocatalyst. The method of claim 7, wherein
A production method of putrescine, which produces putrescine from ornithine. The method of claim 8,
A process for producing putrescine, using a buffer of 0.3 M to 1 M.

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