CN108251405A - Complex enzyme and additive and their application and the method for removing mycotoxin - Google Patents

Abstract

The present invention relates to mycotoxins to remove field, discloses a kind of complex enzyme and additive and their application and the method for removing mycotoxin.Specifically, being related to a kind of complex enzyme, which contains amidase and esterase, which can remove mycotoxin, particularly fumonisin, ochratoxin and T2 toxin；Additive containing the complex enzyme and they in application of the removing mycotoxin particularly in fumonisin, ochratoxin and T2 toxin and the method for removing mycotoxin.Pass through above-mentioned technical proposal, as above amidase and esterase are applied in combination, 2 toxin of Ochratoxin A, fumonisin and T can be removed simultaneously, and removal efficiency greatly improves than a kind of efficiency of enzyme of single use, and also is able to a certain extent remove vomitoxin, aflatoxin, zearalenone toxin.

Description

Complex enzyme and additive and their application and the method for removing mycotoxin

Technical field

The present invention relates to mycotoxins to remove field, and in particular, to a kind of complex enzyme, the addition containing the complex enzyme The application and a kind of method for removing mycotoxin of agent, the complex enzyme and/or additive in mycotoxin is removed.

Background technology

Mycotoxin is a kind of secondary metabolites that Toxigenic fungi generates in the process in harm, mainly includes aspergillus flavus poison Element, deoxynivalenol (also known as vomitoxin, DON), zearalenone (ZEN) and fumonisin (FUM), reddish brown song Mould toxin and T-2 toxin etc..

In recent years, frequently occurring due to China's extreme climate results in head blight to endanger wheat, based on corn It is very popular, mycetogenetic fumonisin (FUM), ochratoxin and T-2 pollutions aggravate, and pollute exceeded wheat and corn not It is disconnected to increase.The safe utilization of the exceeded grain of toxin is studied, can not only protect the kind grain interests of China peasant, while prevent inflow entrance Safeguard that national food Safety of Food Quality is extremely urgent in grain market.

Removing for mycotoxin, current domestic and international more universal method mainly have physical removal and absorption, chemistry Processing etc..Adsorbent also largely adsorbed feed and food while mycotoxin is adsorbed in trace nutrient, sticked Toxin after soil absorption, can not be decomposed, can cause secondary pollution.

It is one of most important means, but toxin currently to pollute grain to produce ethyl alcohol as fermenting raw materials using mycotoxin It is further concentrated in a large amount of by-product vinasse obtained in fermenting and producing, fumonisin, ochratoxin and T-2 contents are significantly It cannot be utilized more than national limit standard.

Toxin efficiently can be converted into non-toxic products, Environmental Safety by biological eliminating.Have become current mycotoxin to cut Subtract treatment technology approach most promising in technology, it is not only safe and environment-friendly, efficient, but also utilize modern biotechnology It conducts a research and meets very much the development trend of current China's energy-saving and emission-reduction.But at present in terms of biological eliminating, it is difficult to simultaneously efficiently Ground removes a variety of toxin, when using compound enzyme agent, often will appear a kind of removal efficiency of toxin and improves, and its He but has dropped the removal efficiency of toxin.When being related to fumonisin (FUM), ochratoxin and T-2 toxin, due to these three Content of toxins is much lower relative to vomitoxin, aflatoxin and zearalenone toxin, therefore, research relatively also compared with It is few, it is more difficult to realize to efficient removal while them.

Therefore there is an urgent need for develop one kind can energy conservation and environmental protection and a variety of mycotoxins of efficient removal can particularly take off simultaneously The complex enzyme formulation removed.

Invention content

The purpose of the invention is to overcome the disadvantages described above of the prior art, providing one kind can while efficiently remove more The compounding complex enzyme formulation of kind mycotoxin particularly fumonisin, ochratoxin and T2 toxin, contains the complex enzyme formulation Additive, the complex enzyme formulation and/or additive are in removing mycotoxin particularly fumonisin, ochratoxin and T2 poison Application and a kind of method for removing mycotoxin particularly fumonisin, ochratoxin and T2 toxin in element.

To achieve these goals, on the one hand, the present invention provides a kind of complex enzyme, wherein, which contains amide Enzyme and esterase, the complex enzyme can remove mycotoxin, particularly fumonisin, ochratoxin and T2 toxin.

Second aspect, the present invention also provides a kind of additive, wherein, which contains complex enzyme as described above.

The third aspect, the present invention provides complex enzyme as described above and/or additive as described above in removing fungi Application in toxin particularly fumonisin, ochratoxin and T2 toxin.

Fourth aspect, the present invention provides a kind of method for removing mycotoxin, wherein, this method includes：To as above institute The complex enzyme and/or additive as described above stated are contacted with the sample that mycotoxin pollutes, to remove the poison of the fungi in sample Element, the mycotoxin are preferably fumonisin, ochratoxin and T2 toxin.

Through the above technical solutions, amidase as above and esterase are applied in combination, can remove simultaneously Ochratoxin A, Fumonisin and T-2 toxin, and removal efficiency greatly improves, and also be able to certain than a kind of efficiency of enzyme of single use Vomitoxin, aflatoxin, zearalenone toxin are removed in degree.

Other features and advantages of the present invention will be described in detail in subsequent specific embodiment part.

Specific embodiment

The specific embodiment of the present invention is described in detail below.It is it should be understood that described herein specific Embodiment is merely to illustrate and explain the present invention, and is not intended to restrict the invention.

The endpoint of disclosed range and any value are not limited to the accurate range or value herein, these ranges or Value should be understood to comprising the value close to these ranges or value.For numberical range, between the endpoint value of each range, respectively It between the endpoint value of a range and individual point value and can be individually combined with each other between point value and obtain one or more New numberical range, these numberical ranges should be considered as specific open herein.

In a first aspect, the present invention provides a kind of complex enzyme, wherein, which contains amidase and esterase, this is compound Enzyme can remove mycotoxin.

According to the present invention, amidase and esterase are carried out with the use of can be removed to a variety of mycotoxins, but work as When the mycotoxin is related to fumonisin, ochratoxin and T2 toxin, removal efficiency significantly improves, and compared to single Enzyme preparation, the removal efficiency of complex enzyme formulation provided by the invention will greatly promote.

Although amidase and esterase to be used cooperatively and can achieve the object of the present invention, it was found by the inventors of the present invention that By using the amidase with following amino acid sequence, the complex enzyme is for mycotoxin, particularly fumonisin, reddish brown song The removal efficiency of mould toxin and T2 toxin can be promoted further.Specifically, the amidase has：(a)SEQID NO：2 Shown amino acid sequence is (by SEQ ID NO：It is nucleotide sequence coded shown in 1)；(b)SEQ ID NO：Amino shown in 2 One or several in the amino acids residue of the 11st, 89,179 and 239 still have after replacing, lacking or add in acid sequence SEQ ID NO：The amino acid sequence of amino acid sequence activity shown in 2 preferably has by SEQID NOs：3rd, shown in 4 and 5 Nucleotide sequence coded amino acid sequence.More preferably, the amino acid sequence of the amidase is SEQ ID NO：Shown in 2 Amino acid or for by SEQ ID NOs：3rd, the nucleotide sequence coded amino acid sequence shown in 4 and 5.

Although amidase and esterase to be used cooperatively and can achieve the object of the present invention, it was found by the inventors of the present invention that By using the esterase with following amino acid sequence, the complex enzyme is for mycotoxin, particularly fumonisin, Aspergillus ochraceus The removal efficiency of toxin and T2 toxin can be promoted further.Specifically, the esterase have SEQ ID NOs: 6,7,8,9, 10th, the amino acid sequence encoded described in nucleotide sequence shown at least one of 11,12,13,14,15,16 and 17.Preferably, The amino acid sequence of the esterase is at least one of SEQ ID NOs: 6,7,8,9,10,11,12,13,14,15,16 and 17 institutes Show the amino acid sequence encoded described in nucleotide sequence.

Wherein, it after as above amidase and the preferred amino acid sequence of esterase has been understood, can be pushed away according to codon is counter Method be readily available the nucleotide sequence for encoding the amidase and esterase.Simultaneously as there are degeneracies for codon, it can To obtain a plurality of coding same amino acid sequence but the different nucleotide sequence of nucleotide sequence, these should be the present invention's Within protection domain.

20 kinds of amino acid residues of constitutive protein matter, four classes are segmented into according to pendant polar：1st, nonpolar amino acid： Alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), methionine (Met), phenylalanine (Phe), tryptophan (Trp) and proline (Pro)；2nd, the uncharged amino acid of polarity：Glycine (Gly), serine (Ser), threonine (Thr), cysteine (Cys), aspartic acid (Asn), glutamine (Gln) and tyrosine (Tyr)；3rd, band The amino acid of positive charge：Arginine (Arg), lysine (Lys) and histidine (His)；4th, negatively charged amino acid：Asparagus fern ammonia Sour (Asp) and glutamic acid (Glu) (referring to " biochemistry " (second edition) first volume, Shen Tong, Wang Jingyan, the 82-83 pages, high religion Educate publishing house, December nineteen ninety).If it happens the amino acid residue substitution of a classification is belonged in protein, such as is taken by Arg Replace Ile for Lys or by Leu, effect of the residue played in protein domain (for example provides positive charge or formed and dredged The effect of water bag structure) do not change, therefore the stereochemical structure of protein can't be had an impact, therefore still can be real The function of existing albumen.The amino acid residue substitution for belonging to a classification can be happened at any one amino acid of above-mentioned enzyme On resi-dues.

As previously mentioned, enzyme provided by the invention can also be modified or is mutated, derivative protein is obtained.Institute of the present invention It states " derivative protein " to refer to the enzyme with above-mentioned amino acid sequence with the difference on amino acid sequence, it is possibility to have not shadow It rings the difference on the modified forms of sequence or haves both at the same time.These albumen include natural or induction genetic variant.It is described Induction variant can be obtained by various technologies, such as the random mutation of radiation or mutagens generation, can also be by such as fixed The technology of point mutation method or other known molecular biology." the derivative protein " is further included with natural L-form amino acid Residue analog (such as D types amino acid) and with it is non-naturally occurring or synthesis amino acid (such as beta-amino acids, γ- Amino acid etc.) analog.

Modification (not changing primary structure usually, i.e., do not change amino acid sequence) form includes：In vivo or in vitro albumen Chemical derivative form such as acetylation or carboxylated.Modification further include glycosylation, such as those in the synthesis and processing of albumen or Albumen that is glycosylation modified and generating is carried out in further processing step.This modification can be by being exposed to progress sugar by albumen The enzyme (glycosylase or deglycosylation enzyme of such as mammal) of base and complete.Modified forms are further included with phosphorylation amino The sequence of sour residue (such as phosphotyrosine, phosphoserine, phosphothreonine).It further includes and is modified to improve its anti-egg White hydrolysis property or the albumen for optimizing solubility property.And in the present invention, in asparagine (Asn), the silk of the complex enzyme Propylhomoserin (Ser) and threonine (Thr) residue carry out that N- and/or O- is glycosylation modified can further to improve dissolubility at any one And thermal stability.

In order to facilitate purifying, the label that this field can also be used common (such as Poly-Arg, Poly-His, FLAG, At least one of Strep-tag II and c-myc) modification is added to (a) or (b).The label does not interfere with the present invention The activity of the enzyme of offer in actual application, can choose whether addition label according to demand.

In order to further improve removal effect of the enzyme compound formulation to mycotoxin, under preferable case, the amidase and The weight ratio of esterase is 1：(0.01-100), further preferably 1：(0.05-20) is still more preferably 1：(0.1-10), More preferably 1：(0.2-5).

The present inventor has found in the course of the study, when also containing phosphatidyl silk in the enzyme compound formulation of the present invention In the case of propylhomoserin and/or Zn-ef ficiency, the removal efficiency of mycotoxin can be promoted further.Preferably, with described compound On the basis of the total weight of enzyme preparation, the content of the phosphatidylserine and/or Zn-ef ficiency is 1-30 weight %, more preferably 5- 10 weight %.

In the present invention, the amidase and esterase can be prepared by existing method, for example, by that will encode correlation The nucleotide sequence of amino acid sequence is cloned into carrier, then is transferred in host, then by conventional method from the place after proliferation The isolated related nucleotide sequence of chief cell, can also be commercially available, the phosphatidylserine and/or Zn-ef ficiency are equal It can be commercially available, can also be obtained by other regular approach.

According to the present invention, in order to further enhance the detoxification efficiency of the complex enzyme, the complex enzyme also containing a kind of or A variety of other enzymes；One or more other enzymes can be selected from but be not limited to：Aflatoxin detoxifying enzymes, Aspergillus ochraceus Toxin lactonase, fumonisin Carboxylesterase, fumonisin aminopherase, amino polyol amine oxidase, deoxidation melon wither sickle Mykol epoxide hydrolase, carboxypeptidase, aspergillus niger aspartic protease PEPAa, PEPAb, PEPAc and PEPAd, elastic egg White enzyme, aminopeptidase, pepsin or pepsin sample protease, trypsase or trypsin like proteases, bacterioprotein Enzyme, be related to starch metabolism, fiber removal, lipid-metabolism enzyme, be related to glycogen metabolism protein or enzyme, amylase, Arab Carbohydrase, arabinofuranosidase, catalase, cellulase, chitinase, renin, cutinase, DNA Enzyme, epimerase, esterase, galactosidase, dextranase, glucan lyase, endoglucanase, glucoamylase, Portugal Grape carbohydrate oxidase ,-glucosidase, including β-glucosyl enzym, glycuronidase, hemicellulase, hexoxidase, hydrolase, Invertase, isomerase, lipolytic enzyme, laccase, lyases, mannosidase, oxidizing ferment, oxidoreducing enzyme, pectate lyase, pectin Acetylesterase, pectin remove polymerase, pectinesterase, pectin decomposing enzyme, peroxidase, phenol oxidase, phytase, poly half Lactobionic acid enzyme, protease, sandlwood-galacturonic acid enzyme, African hesperidium element, transferase, transport protein, turn ribalgilase Glutaminase, zytase, hexoxidase, acid phosphatase and combination thereof.

It in the present invention, uses for convenience, under preferable case, each component of the enzyme compound formulation may be mixed together Packaging, can also respective independent packaging, then mixed again when in use.

The present invention also provides the preparation method of above-mentioned enzyme compound formulation, this method includes：It will according to aforementioned weight ratio Each ingredient is uniformly mixed.

According to the present invention, the fumonisin can include its each Type of toxin, for example, FA1, FA2, FB1, FB2, FB3, FB4, FC1, FC2, FC3, FC4 and FP1, wherein, most preferably fumonisin FB1.The ochratoxin can also include it Each Type of toxin, for example, ochratoxin A and ochratoxin B.

Second aspect, the invention also discloses a kind of additive, wherein, which contains complex enzyme as described above.

In situations where it is preferred, the additive is using complex enzyme provided by the invention as active constituent.In the addition In agent, the content of the complex enzyme is 0.001-10g/kg, more preferably preferably 0.01-8g/kg, 0.1-5g/kg.In addition, It can also be containing well known to a person skilled in the art solvent (such as glycerine, carbohydrate and protease inhibitors egg in the additive White protective agent), agonist etc..

According to the present invention, also containing physiologically acceptable carrier in the additive, wherein, it is described physiologically to connect The carrier received is selected from least one of following substance：Maltodextrin, lime stone (calcium carbonate), cyclodextrin, wheat, wheat bran or Wheat component, rice or rice bran, sucrose, starch, Na₂SO₄, talcum powder and PVA and their mixture.

It will be apparent for a person skilled in the art that can these additives be added to pollution has the feed of mycotoxin And/or grain and oil material is to reduce existing endotoxin level.

The Feed Material of the present invention can include：A) cereal, for example, granule cereal (such as wheat, barley, naked barley, oat with And combination thereof) and/or big grain cereal such as maize or sorghum；B) by-product from cereal, for example, it is corn protein powder, dry Vinasse and soluble matter (DDGS), wheat bran, sizing, wheat wheat-middlings, rice bran, rice husk, oat shell, palm kernel and citrus pulp；c) Ensilage；D) protein derived from following source：Such as soybean, sunflower, peanut, lupin, pea, broad bean, cotton, card Nola, fish meal, dry plasma albumen, meat and bone meal, potato protein, whey, copra, sesame；E) from plant and animal source The oil ＆ fat of acquisition；F) minerals and vitamins.

The grain and oil of the present invention refer to the grains such as cereal, beans and oil plant and its general designation of processing finished product and semi-finished product, special Do not refer to the edible product of the mankind.For example, the grain and oil can be the edible grain and oil production of the common mankind in this field Product, specifically, the grain and oil can include in cereal and its agricultural and sideline product, oil ＆ fat product, drinks, milk and its product etc. It is at least one.

It will be apparent to one skilled in the art that feed according to the present invention or additive can also include other groups Divide such as stabilizer and/or incremental agent and/or enzyme.

Preferably, the method for preparing additive according to the present invention includes mixing step, which includes mixing such as The upper complex enzyme, carrier optionally physiologically acceptable at least one, solvent, agonist, stabilizer, incremental agent or enzyme Class is mixed together.

It is apparent to the skilled person that, as precautionary step, any feed and/or grain and oil can be added an additive to In material.

For example, when the additive is used for the feed of cultivated animals, in the additive also containing bacillus licheniformis, Bacillus subtilis, bifidobacterium bifidum, enterococcus faecalis, enterococcus faecium, lactoenterococcus, lactobacillus acidophilus, Lactobacillus casei, German-style Lactobacillus lactate subspecies, lactobacillus plantarum, Pediococcus acidilactici, Pediococcus pentosaceus, candida utili, baby's bifid bar Bacterium, bifidobacterium longum, bifidobacterium breve, bifidobacterium adolescentis, streptococcus thermophilus, lactobacillus reuteri, animal bifidobacteria, rice Aspergillus, bacillus lentus, bacillus pumilus, lactobacillus cellobiosas, lactobacillus fermenti and Lactobacillus delbrueckii At least one of subspecies.

For example, when the additive is used for ensilage or ox feed, also containing production propionic acid propionic acid in the additive At least one of bacillus, lactobacillus buchneri and Lactobacillus paracasei.

For example, when the additive be used for poultry, growing-finishing pig and aquiculture animal feed when, the feed or Also contain bacillus coagulans and/or Brevibacillus laterosporus in additive.

Based on description as above, the present invention also provides a kind of grain and oil or feed, wherein, the grain and oil or feed contain above-mentioned Additive.

The third aspect is true in removing the present invention also provides complex enzyme as described above and/or additive as described above Application in verticillium toxin particularly fumonisin, ochratoxin and T2 toxin.

Fourth aspect, the present invention also provides a kind of method for removing mycotoxin, wherein, this method includes：It will as above The complex enzyme and/or additive is contacted with the sample that mycotoxin pollutes, described true to remove the mycotoxin in sample Verticillium toxin is preferably fumonisin, ochratoxin and T2 toxin.

According to the present invention, contact each enzyme component preferably in the complex enzyme can play under conditions of enzymatic activity into Row.Wherein, term " condition for playing enzymatic activity " refers to meet the conditions such as temperature and the pH of enzymatic activity performance.

According to the present invention, the sample of the mycotoxin pollution can include any sample containing mycotoxin, for example, Can be grain and oil and/or feed etc..It is described compound on the basis of the total weight of the sample of mycotoxin pollution under preferable case The dosage of enzyme preparation is is at least 0.1ppm, more preferably preferably 0.1-15ppm, further preferably 1-10ppm, 4- 8ppm.In the present invention, when the sample of mycotoxin pollution is liquid, that " ppm " is represented is " μ g/mL "；Work as vomitoxin When the sample of pollution is solid, that " ppm " is represented is " μ g/g ".

In following embodiment and comparative example, ochratoxin A, fumonisin FB1 and T2 toxin standard items are purchased from Sigma Company；Phosphatidylserine is purchased from upper Hisense's sail bio tech ltd.

Fumonisin FB1 is examined by high performance liquid chromatography according to the method in GB 5009.240-2016 standards It surveys, vomitoxin is detected according to the method in GB/T 30956-2014 standards, according to GB/T 23501-2009 standards In method T2 toxin is detected；

Toxin degradation rate %=(before reaction in sample after quality-reaction of toxin in sample toxin quality)/reaction before Quality × 100% of toxin in sample.

Embodiment

The present invention will be described in detail by way of examples below, but is not intended to limit the present invention.Following preparation example, In embodiment and comparative example, unless otherwise instructed, each material is commercially available, and each method is the conventional method of this field.

Preparation example 1

This preparation example is used for the preparation for illustrating amidase provided by the invention

(1) acquisition of gene

(precious bioengineering (Dalian) Co., Ltd of commission, similarly hereinafter) following nucleotide is synthesized by artificial chemistry synthetic method Segment；In SEQ ID NO：Addition Nde I restriction enzyme sites, 3 ' ends after 5 ' end initiation codon ATG of the nucleotide sequence shown in 1 Add Xho I restriction enzyme sites and terminator codon TAG.

(2) structure of recombinant plasmid

(there is His labels, purchase to PET30a plasmids using restriction enzyme Nde I and Xho I (being purchased from NEB companies) In Invitrogen companies of the U.S.) double digestion is carried out, in 37 DEG C of water-bath digestion 4h, digestion system (50 μ L) is as follows：

By digestion products into after row agarose gel electrophoresis, purifying recycling.Then, (it is purchased from Takara using T4 ligases Company) reaction is attached, 4h is connected at room temperature, obtains recombinant plasmid.The linked system (10 μ L) used is as follows：

(3) acquisition of recombinant bacterial strain

Turn DH5 α competence using the recombinant plasmid electricity obtained by step (2) on Bio-Rad Gene Pulse electroporations Cell (is purchased from Takara companies), and electric conversion condition is：Voltage 1500V, 25 μ F of capacitance, 200 Ω of resistance, transformation time is with DNA The difference of sample and change, and provided automatically by instrument, usually in the range of 3.5-4s.Then, the cell after conversion is coated with It is screened on the LB solid plates containing kanamycins, is inverted culture 2 days in 37 DEG C, picking positive bacterium colony is inoculated in LB liquid In body culture medium, to obtain recombinant bacterial strain.

Then make with method known in this field to extract plasmid, then entrust precious bioengineering (Dalian) limited public affairs Department is sequenced, the results show that gene as above is successfully transformed into Escherichia coli, shows the recombinant bacterial strain of the present invention It builds successfully.

(4) preparation of amidase

Obtained recombinant bacterial strain is inoculated in LB fluid nutrient mediums, and (beef extract 5g, peptone 10g, sodium chloride 5g, moisturizing is extremely 1000mL, 115 DEG C of sterilizing 20min are spare, pH 7) in, 3 days are cultivated in 37 DEG C to OD₆₀₀Between 2 and 6, bacterium is collected by centrifugation in value Body, with phosphate buffer (PBS, 135mM NaCl, 2.7mM KCl, 1.5mM KH₂PO₄, 8mM K₂HPO₄, pH 7.2) and solution After suspension, ultrasonic disruption, centrifuging and taking supernatant obtains crude enzyme liquid.

Crude enzyme liquid is placed in the ammonium sulfate powder for, being slowly added to grind on ice, it is stirring while adding, it is added to ammonium sulfate saturation Until.Left and right for 24 hours is stood at 4 DEG C, 12000r/min centrifugation 50min abandon supernatant, dissolved and precipitated with a small amount of PBS (pH 7.2).It will The precipitation dialysis of PBS dissolvings, removes ammonium sulfate, is resuspended in buffer solution (pH 7.4,50mM NaCl, imidazoles containing 10mM).According to The recombinase given expression to contains His labels, affinitive layer purification is carried out using Ni columns, after 1mL/min balances Ni columns, with flow 0.5mL/min is directly by the crude enzyme liquid loading of resuspension；It is continuing with buffer solution (pH7.4,50mM NaCl, imidazoles containing 10mM) 1mL/min elutes unadsorbed or absorption non-specific foreign protein；With buffer solution (pH 7.4,50mM NaCl, 500mM imidazoles) Destination protein is collected in elution, to obtain the amide enzyme solution of purifying, and is freeze-dried and is obtained amidase dry powder X1.

Preparation example 2-4

Preparation example 2-4 is used for the preparation for illustrating amidase provided by the invention

The preparation of amidase X2-X4 is carried out respectively according to the method for preparation example 1, unlike, preparation example 2-4 is used respectively SEQ ID NO：3、SEQ ID NO：4 and SEQ ID NO：5 replace SEQ ID NO：1.

Preparation example 5-16

Preparation example 5-16 is used for the preparation method for illustrating esterase provided by the invention

The preparation of esterase Z1-Z12 is carried out respectively according to the method for preparation example 1, unlike, preparation example 5-16 is used respectively SEQ ID NO：6、SEQ ID NO：7、SEQ ID NO：8、SEQ ID NO：9、SEQ ID NO：10、SEQ ID NO：11、SEQ ID NO：12、SEQ ID NO：13、SEQ ID NO：14、SEQ ID NO：15、SEQ ID NO：16 and SEQ ID NO：17 replace Change SEQ ID NO：1.

Embodiment 1-14, comparative example 1-3

For illustrating that complex enzyme provided by the invention makees the removing of fumonisin FB1, ochratoxin A and T2 toxin With.

The preparation of complex enzyme formulation is carried out according to the proportioning of each embodiment and comparative example in table 1 using physiological saline so that In obtained mixed liquor, a concentration of 100ng/ml of complex enzyme, the concentration point of fumonisin FB1, ochratoxin A and T2 toxin It Wei not 1000ppb.For reaction temperature to be respectively 37 DEG C, the pH value of reaction is respectively 5, and 20 μ L reaction products are examined after reacting 30min The residual of fumonisin FB1, ochratoxin A and T2 toxin is surveyed, and calculates degradation rate.The results are shown in Table 2.

Table 1

	Amidase	Esterase	Phosphatidylserine	Zinc
					Embodiment 1	X1 50ng/ml	Z1 50ng/ml	5ng/ml	---
Embodiment 2	X2 80ng/ml	Z2 20ng/ml	---	8ng/ml
					Embodiment 3	X3 20ng/ml	Z3 80ng/ml	5ng/ml	5ng/ml
Embodiment 4	X1 50ng/ml	Z1 50ng/ml	---	---
					Embodiment 5	X1 50ng/ml	Z4 50ng/ml	5ng/ml	---
Embodiment 6	X1 50ng/ml	Z5 50ng/ml	5ng/ml	---
					Embodiment 7	X1 50ng/ml	Z6 50ng/ml	5ng/ml	---
Embodiment 8	X1 50ng/ml	Z7 50ng/ml	5ng/ml	---
					Embodiment 9	X1 50ng/ml	Z8 50ng/ml	5ng/ml	---
Embodiment 10	X1 50ng/ml	Z9 50ng/ml	5ng/ml	---
					Embodiment 11	X1 50ng/ml	Z10 50ng/ml	5ng/ml	---
Embodiment 12	X1 50ng/ml	Z11 50ng/ml	5ng/ml	---
					Embodiment 13	X1 50ng/ml	Z12 50ng/ml	5ng/ml	---
Embodiment 14	X4 50ng/ml	Z1 50ng/ml	5ng/ml	---
					Comparative example 1	---	---	5ng/ml	---
Comparative example 2	X1 100ng/ml	---	5ng/ml	---
					Comparative example 3	---	Z1 100ng/ml	5ng/ml	---

Table 2

Amidase and esterase are combined use it can be seen from the data of upper table 2, can efficiently remove volt horse Toxin and ochratoxin, and most T2 toxin can also be removed.And work as in the complex enzyme and add phosphide When acyl serine and/or zinc, the removal efficiency of toxin can be promoted further.

The preferred embodiment of the present invention has been described above in detail, still, during present invention is not limited to the embodiments described above Detail, within the scope of the technical concept of the present invention, a variety of simple variants can be carried out to technical scheme of the present invention, this A little simple variants all belong to the scope of protection of the present invention.

It is further to note that specific technical features described in the above specific embodiments, in not lance In the case of shield, it can be combined by any suitable means.In order to avoid unnecessary repetition, the present invention to it is various can The combination of energy no longer separately illustrates.

In addition, various embodiments of the present invention can be combined randomly, as long as it is without prejudice to originally The thought of invention, it should also be regarded as the disclosure of the present invention.

SEQUENCE LISTING

<110>Cofco Nutrition And Health Research Institute Co., Ltd.

Cofco Group Co., Ltd.

<120>Complex enzyme and additive and their application and the method for removing mycotoxin

<130> I40389COF

<160> 17

<170> PatentIn version 3.3

<210> 1

<211> 1443

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 1

atggtccgcc gaattgcttc agctacacct cgcgtgcaat cgcccatgtc gccattgggc 60

acaacatact gcgtccgtcc taatcctgtt tcactgaatc ttcaaagaag acctctcgtg 120

atcgcatcaa cagacgaggc caaggtcact ataatatatg ccggactatt aatccctggc 180

gacggagaac ctctgcgcaa tgctgcccta gtcatcagcg ataagatcat cgcgttcgtt 240

ggatccgaag ccgacatccc taaggactac ctccggtcca cgcagtctac tcatcgtgtc 300

cccgtgctca tgcctggttt gtgggattgc gacatgcatt ttggcgggga tgacgattat 360

tacaacgatt atacatctgg tctggccact catccagcat catcaggtgc tcgactagcc 420

cgtggttgct gggaagcatt gcagaatggg tatacatcct accgcgacct agccggatac 480

gggtgcgagg tcgcaaaggc gatcaatgat ggcactatcg ttggtccaaa cgtgcattcg 540

tctggcgctg cactcagtca gacagctgga cacggcgata tcttcgctct tccagcaggc 600

gaagtactgg ggagttatgg agtaatgaac ccacgccctg ggtactgggg ggcagggccg 660

ctatgtatcg ccgatggcgt agaggaggtc cgacgagcag tgaggttgca gatcatgcgc 720

ggtgcaaagg ttatcaaagt gatggcctct gggggtgtca tgtcgcgaga cgataatccc 780

aactttgcac agttctctcc agaagaactg aaggtgatag tggaagaggc ggctcgacag 840

aaccggatcg tttctgcaca tgtgcatggc aaggcgggga ttatggctgc tatcaaagca 900

ggctgcaaga gtctggagca tgtgtcttat gctgacgagg aggtctggga gctcatgaaa 960

gagaagggaa ttttgtatgt ggccacacgc tcggttattg aaatctttct ggctagtaat 1020

ggagaggggt tggtgaaaga gtcgtgggcc aagttgcagg cccttgccga ttcgcatttg 1080

aaagcttatc agggagctat taaggcgggt gttaccattg cgttgggaac ggataccgcc 1140

cccggtggtc ctaccgcact tgagttgcag tttgccgtcg agagaggagg tatgacgccg 1200

ttggaggcca tcaaagccgc aactgcgaac gctcccctgt cagttggtcc acaagcaccg 1260

ttgacgggtc agcttcgcga ggggtatgag gcagatgtga ttgcgttgga ggagaatcca 1320

ttggaggaca tcaaagtctt tcaggagccg aaggcagtta cccacgtctg gaagggaggg 1380

aaactgttca aaggtccagg tattggtccg tggggagaag atgcacgtaa tccttttctg 1440

tag 1443

<210> 2

<211> 480

<212> PRT

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 2

Met Val Arg Arg Ile Ala Ser Ala Thr Pro Arg Val Gln Ser Pro Met

1 5 10 15

Ser Pro Leu Gly Thr Thr Tyr Cys Val Arg Pro Asn Pro Val Ser Leu

20 25 30

Asn Leu Gln Arg Arg Pro Leu Val Ile Ala Ser Thr Asp Glu Ala Lys

35 40 45

Val Thr Ile Ile Tyr Ala Gly Leu Leu Ile Pro Gly Asp Gly Glu Pro

50 55 60

Leu Arg Asn Ala Ala Leu Val Ile Ser Asp Lys Ile Ile Ala Phe Val

65 70 75 80

Gly Ser Glu Ala Asp Ile Pro Lys Asp Tyr Leu Arg Ser Thr Gln Ser

85 90 95

Thr His Arg Val Pro Val Leu Met Pro Gly Leu Trp Asp Cys Asp Met

100 105 110

His Phe Gly Gly Asp Asp Asp Tyr Tyr Asn Asp Tyr Thr Ser Gly Leu

115 120 125

Ala Thr His Pro Ala Ser Ser Gly Ala Arg Leu Ala Arg Gly Cys Trp

130 135 140

Glu Ala Leu Gln Asn Gly Tyr Thr Ser Tyr Arg Asp Leu Ala Gly Tyr

145 150 155 160

Gly Cys Glu Val Ala Lys Ala Ile Asn Asp Gly Thr Ile Val Gly Pro

165 170 175

Asn Val His Ser Ser Gly Ala Ala Leu Ser Gln Thr Ala Gly His Gly

180 185 190

Asp Ile Phe Ala Leu Pro Ala Gly Glu Val Leu Gly Ser Tyr Gly Val

195 200 205

Met Asn Pro Arg Pro Gly Tyr Trp Gly Ala Gly Pro Leu Cys Ile Ala

210 215 220

Asp Gly Val Glu Glu Val Arg Arg Ala Val Arg Leu Gln Ile Met Arg

225 230 235 240

Gly Ala Lys Val Ile Lys Val Met Ala Ser Gly Gly Val Met Ser Arg

245 250 255

Asp Asp Asn Pro Asn Phe Ala Gln Phe Ser Pro Glu Glu Leu Lys Val

260 265 270

Ile Val Glu Glu Ala Ala Arg Gln Asn Arg Ile Val Ser Ala His Val

275 280 285

His Gly Lys Ala Gly Ile Met Ala Ala Ile Lys Ala Gly Cys Lys Ser

290 295 300

Leu Glu His Val Ser Tyr Ala Asp Glu Glu Val Trp Glu Leu Met Lys

305 310 315 320

Glu Lys Gly Ile Leu Tyr Val Ala Thr Arg Ser Val Ile Glu Ile Phe

325 330 335

Leu Ala Ser Asn Gly Glu Gly Leu Val Lys Glu Ser Trp Ala Lys Leu

340 345 350

Gln Ala Leu Ala Asp Ser His Leu Lys Ala Tyr Gln Gly Ala Ile Lys

355 360 365

Ala Gly Val Thr Ile Ala Leu Gly Thr Asp Thr Ala Pro Gly Gly Pro

370 375 380

Thr Ala Leu Glu Leu Gln Phe Ala Val Glu Arg Gly Gly Met Thr Pro

385 390 395 400

Leu Glu Ala Ile Lys Ala Ala Thr Ala Asn Ala Pro Leu Ser Val Gly

405 410 415

Pro Gln Ala Pro Leu Thr Gly Gln Leu Arg Glu Gly Tyr Glu Ala Asp

420 425 430

Val Ile Ala Leu Glu Glu Asn Pro Leu Glu Asp Ile Lys Val Phe Gln

435 440 445

Glu Pro Lys Ala Val Thr His Val Trp Lys Gly Gly Lys Leu Phe Lys

450 455 460

Gly Pro Gly Ile Gly Pro Trp Gly Glu Asp Ala Arg Asn Pro Phe Leu

465 470 475 480

<210> 3

<211> 1443

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 3

atggtccgcc gaattgcttc agctacacct cacgtgcaat cgcccatgtc gccattgggc 60

acaacatact gcgtccgtcc taatcctgtt tcactgaatc ttcaaagaag acctctcgtg 120

atcgcatcaa cagacgaggc caaggtcact ataatatatg ccggactatt aatccctggc 180

gacggagaac ctctgcgcaa tgctgcccta gtcatcagcg ataagatcat cgcgttcgtt 240

ggatccgaag ccgacatccc taaggactac ctccggtcca cgcagtctac tcatcgtgtc 300

cccgtgctca tgcctggttt gtgggattgc gacatgcatt ttggcgggga tgacgattat 360

tacaacgatt atacatctgg tctggccact catccagcat catcaggtgc tcgactagcc 420

cgtggttgct gggaagcatt gcagaatggg tatacatcct accgcgacct agccggatac 480

gggtgcgagg tcgcaaaggc gatcaatgat ggcactatcg ttggtccaaa cgtgcattcg 540

tctggcgctg cactcagtca gacagctgga cacggcgata tcttcgctct tccagcaggc 600

gaagtactgg ggagttatgg agtaatgaac ccacgccctg ggtactgggg ggcagggccg 660

ctatgtatcg ccgatggcgt agaggaggtc cgacgagcag tgaggttgca gatctttcgc 720

ggtgcaaagg ttatcaaagt gatggcctct gggggtgtca tgtcgcgaga cgataatccc 780

aactttgcac agttctctcc agaagaactg aaggtgatag tggaagaggc ggctcgacag 840

aaccggatcg tttctgcaca tgtgcatggc aaggcgggga ttatggctgc tatcaaagca 900

ggctgcaaga gtctggagca tgtgtcttat gctgacgagg aggtctggga gctcatgaaa 960

gagaagggaa ttttgtatgt ggccacacgc tcggttattg aaatctttct ggctagtaat 1020

ggagaggggt tggtgaaaga gtcgtgggcc aagttgcagg cccttgccga ttcgcatttg 1080

aaagcttatc agggagctat taaggcgggt gttaccattg cgttgggaac ggataccgcc 1140

cccggtggtc ctaccgcact tgagttgcag tttgccgtcg agagaggagg tatgacgccg 1200

ttggaggcca tcaaagccgc aactgcgaac gctcccctgt cagttggtcc acaagcaccg 1260

ttgacgggtc agcttcgcga ggggtatgag gcagatgtga ttgcgttgga ggagaatcca 1320

ttggaggaca tcaaagtctt tcaggagccg aaggcagtta cccacgtctg gaagggaggg 1380

aaactgttca aaggtccagg tattggtccg tggggagaag atgcacgtaa tccttttctg 1440

tag 1443

<210> 4

<211> 1440

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 4

atggtccgcc gaattgcttc agctacacct gtgcaatcgc ccatgtcgcc attgggcaca 60

acatactgcg tccgtcctaa tcctgtttca ctgaatcttc aaagaagacc tctcgtgatc 120

gcatcaacag acgaggccaa ggtcactata atatatgccg gactattaat ccctggcgac 180

ggagaacctc tgcgcaatgc tgccctagtc atcagcgata agatcatcgc gttcgttgga 240

tccgaagccg acatccctaa ggactacctc cggtccacgc agtctactca tcgtgtcccc 300

gtgctcatgc ctggtttgtg ggattgcgac atgcattttg gcggggatga cgattattac 360

aacgattata catctggtct ggccactcat ccagcatcat caggtgctcg actagcccgt 420

ggttgctggg aagcattgca gaatgggtat acatcctacc gcgacctagc cggatacggg 480

tgcgaggtcg caaaggcgat caatgatggc actatcgttg gtccaaacgt gcattcgtct 540

ggcgctgcac tcagtcagac agctggacac ggcgatatct tcgctcttcc agcaggcgaa 600

gtactgggga gttatggagt aatgaaccca cgccctgggt actggggggc agggccgcta 660

tgtatcgccg atggcgtaga ggaggtccga cgagcagtga ggttgcagat ctttcgcggt 720

gcaaaggtta tcaaagtgat ggcctctggg ggtgtcatgt cgcgagacga taatcccaac 780

tttgcacagt tctctccaga agaactgaag gtgatagtgg aagaggcggc tcgacagaac 840

cggatcgttt ctgcacatgt gcatggcaag gcggggatta tggctgctat caaagcaggc 900

tgcaagagtc tggagcatgt gtcttatgct gacgaggagg tctgggagct catgaaagag 960

aagggaattt tgtatgtggc cacacgctcg gttattgaaa tctttctggc tagtaatgga 1020

gaggggttgg tgaaagagtc gtgggccaag ttgcaggccc ttgccgattc gcatttgaaa 1080

gcttatcagg gagctattaa ggcgggtgtt accattgcgt tgggaacgga taccgccccc 1140

ggtggtccta ccgcacttga gttgcagttt gccgtcgaga gaggaggtat gacgccgttg 1200

gaggccatca aagccgcaac tgcgaacgct cccctgtcag ttggtccaca agcaccgttg 1260

acgggtcagc ttcgcgaggg gtatgaggca gatgtgattg cgttggagga gaatccattg 1320

gaggacatca aagtctttca ggagccgaag gcagttaccc acgtctggaa gggagggaaa 1380

ctgttcaaag gtccaggtat tggtccgtgg ggagaagatg cacgtaatcc ttttctgtag 1440

<210> 5

<211> 1446

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 5

atggtccgcc gaattgcttc agctacacct cgcgtgcaat cgcccatgtc gccattgggc 60

acaacatact gcgtccgtcc taatcctgtt tcactgaatc ttcaaagaag acctctcgtg 120

atcgcatcaa cagacgaggc caaggtcact ataatatatg ccggactatt aatccctggc 180

gacggagaac ctctgcgcaa tgctgcccta gtcatcagcg ataagatcat cgcgttcgtt 240

ggatccgaag ccgacatccc taaggactac ctccggtcca cgcagtctac tcatcgtgtc 300

cccgtgctca tgcctggttt gtgggattgc gacatgcatt ttggcgggga tgacgattat 360

tacaacgatt atacatctgg tctggccact catccagcat catcaggtgc tcgactagcc 420

cgtggttgct gggaagcatt gcagaatggg tatacatcct accgcgacct agccggatac 480

gggtgcgagg tcgcaaaggc gatcaatgat ggcactatcg ttggtccaaa cgtgcatcat 540

tcgtctggcg ctgcactcag tcagacagct ggacacggcg atatcttcgc tcttccagca 600

ggcgaagtac tggggagtta tggagtaatg aacccacgcc ctgggtactg gggggcaggg 660

ccgctatgta tcgccgatgg cgtagaggag gtccgacgag cagtgaggtt gcagatcatg 720

cgcggtgcaa aggttatcaa agtgatggcc tctgggggtg tcatgtcgcg agacgataat 780

cccaactttg cacagttctc tccagaagaa ctgaaggtga tagtggaaga ggcggctcga 840

cagaaccgga tcgtttctgc acatgtgcat ggcaaggcgg ggattatggc tgctatcaaa 900

gcaggctgca agagtctgga gcatgtgtct tatgctgacg aggaggtctg ggagctcatg 960

aaagagaagg gaattttgta tgtggccaca cgctcggtta ttgaaatctt tctggctagt 1020

aatggagagg ggttggtgaa agagtcgtgg gccaagttgc aggcccttgc cgattcgcat 1080

ttgaaagctt atcagggagc tattaaggcg ggtgttacca ttgcgttggg aacggatacc 1140

gcccccggtg gtcctaccgc acttgagttg cagtttgccg tcgagagagg aggtatgacg 1200

ccgttggagg ccatcaaagc cgcaactgcg aacgctcccc tgtcagttgg tccacaagca 1260

ccgttgacgg gtcagcttcg cgaggggtat gaggcagatg tgattgcgtt ggaggagaat 1320

ccattggagg acatcaaagt ctttcaggag ccgaaggcag ttacccacgt ctggaaggga 1380

gggaaactgt tcaaaggtcc aggtattggt ccgtggggag aagatgcacg taatcctttt 1440

ctgtag 1446

<210> 6

<211> 1179

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 6

atgcggaacg tcagcgacaa ggcgccgccc cacgagacgc tcaccgtagt cgtcgcggca 60

atgatcgttg gcacggccgc cttgatggtg cttggaatac agcccatcct tctcggcgcc 120

cttgtagagg aggggcgtat tcccgccgag gggttgggat cggcggcaac ggtggaaata 180

ctggcgatcg cggcgggaac atgcatcgga cccgttctta tgaagacggg atatctgcgg 240

gcgaaatgcg cggcactctg cttaatgctc gccgcaatca acttcggatt gacgttgccg 300

ggtttcgatt tgcccatcgt ggcttgccga gcggcagcgg gagccctgga aggtctttcg 360

ctcagcgcgg cgatcctgat catgactcat aatcggcggc cggaccggct gagcggaata 420

tttctgggcg cgcagacgat accgcaggta atatctgctt atttgctccc gacggagatt 480

attccgcgct gggggagcgc aggcggcttc acgatcctgg gcattctcgc ggcgatcgcc 540

gcgatcgcgg ctctgtgcct cgtcgatcgc gttgagctcg atccgacgac cgttaacgac 600

gacttgcagt ggtcacccgc ggcgatcgtc atttcgatgg cggcattcgt tcaattctcg 660

ggggtcggtg ccgcatggag ctatctggag cgactggctg cgcagcacgg attttcggga 720

gaaacgatcg gtatcgccat ttccgggagt ttgctttgcc aggtaggcgg ggcttggctg 780

gccgcttgga tcggtgggcg ggtcggatat cgcttcgcct taatcgctgg gagcctgctt 840

caggcgggca acgtgatcgc attggcggtg gccgatcagc caagctggtt tatttccgct 900

tcctgtgctt tcggcctgtt ctggttggcg atgcagccct tccaaatccg cttcgcgatc 960

gcgatagata acagccggca gcttgctgta ctgctgacgc cgatcgccct cgtcgggttg 1020

agcgcggggc ccttgttgct ctctcgcttt gccggggcga ccgacttgcg ctggatcttt 1080

gtggggagtt cgaccttgtt gctggccagc gcgcttctgt atctttgcgc ttctctgttt 1140

caaccgcgcg gaaaggtgat cgctgaaacg gtggacgta 1179

<210> 7

<211> 651

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 7

atgacatcgc aggtcaagct tcgtagcgcg gcaaagcggc cgcgcagtcc taaaagcgag 60

cgaggtcttg ctcgttacga gtccttgctt gatgcgaccg acaggctgtt ggtcgatcta 120

gaccccgatc aggtcggtct ctatcagatt gcagaggaag cgggtgcctc accgtcgtcc 180

gtctatcatt tctttccgac caaggaagtg gctcatctcg ctctgatgcg ccgctatctg 240

gaggggctcc ggaatctcga cgcgatggaa gtcgacatcg gccagctcga aagctggcag 300

gacctgatga agttggatca gatcagggcg cgagactatt ataatagcca cccgcccgcc 360

ctcaagcttc tgttcggcgg atatggcggg gtcgaggcca gaaagcttga cgagcgatac 420

tccgaggaaa tcgtgagctc catgtatggc agatacaacg gcattttcca tatgccgcaa 480

atggagaatg aggctctcat gttcacgatc tgcttcgcaa ttctcgacgc ggtatgggcc 540

gtctcctttc gccggttcgg tgaaattacg tcggattttc ttcgggaggg gcaagcggct 600

tgcattgcct attgccgaca ctatctgccc gagcgaacgc catcagcgtg a 651

<210> 8

<211> 945

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 8

gtggccagca agttcaactg tgagttactc gatctgcgat catttgttgc ggtgtatgaa 60

acgcgaagtt ttagccacgc cgcgcggctt ctgaatcaat cgcagcccgc gctcagccgg 120

agaatccagc gcctcgagag tctcgtgggc ggtccgttgt tcgagcggac cagtcggtcg 180

cttgccgaaa cggcgctcgg caaagagttg ctcccggtcg cccaccgagc gttggaactt 240

gtcgatacgt cgctgtttgc gtcgcccaat gtccgggagt tccgctggac agacatcacg 300

attgcctgtg tacagaccgc cgccttccat gttctcccgc gagctgcgcg cttgtacatg 360

gatcaaaatc cgagggtccg actccgcatc cttgacgtgc cggcggtcga ggctgcggac 420

ctggttgcga gcggcgaggc ggagttcggc atcagcattg agagcctgtt gccatcaagc 480

ctgcggttcg atgcgctcca cgaggacccg ttcggcctgg catgccaccg aagccatccg 540

ctggcgtcgc tcgagatcct tgaatggacg caattgaaag gtgaaagcct gatcgccgtt 600

caccgtgcga gccggaaccg cacgttgctc gatgccgaac tcgcgcgcaa caatatcgcg 660

ctggaatggc ggtatgaggt cgcgcatctg acgacggcgc tgggattgat cgatgcgcaa 720

ttgggtgtcg ctgttatgcc ccgcatggtt atgccccgct cgggtcggtc ggaggtcgtc 780

tggcgccccg tcgtcgcgcc ggtcgtccaa cgcacgatcg gcatcgttca gcgccgcacc 840

ggctcgatgc accctgccgc acagcaattg cttgcgcggc tccgcgcggc ctggtcgtcc 900

gccaatctgg gcgacatcgc gtctcgcgaa gatggggcat cgtga 945

<210> 9

<211> 1623

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 9

gtgaaagagc accaatgccg tggcggccgg gcgtcccccg ctgcgcccgc cacgtggctt 60

gcgcggatca gcgtttcccg gggggcctcc gccatcgcct ggaccttcat gcttggcgca 120

actgccattc ccgtggctgc gcaaactgac gatccgaagc tcgttcgtca tacccagtcg 180

ggcgccgtcg agggcgtcga gggcgacgtc gagacttttt tgggaatacc cttcgcggct 240

ccgccggtcg gcgacctgcg atggcggccg ccggctccgc cgagggcgtg ggcgggcacc 300

agggacggcc gccgctttgc gcccgattgc atcgggaacg agcggcttag agaggggagc 360

cgggctgccg ggacgagcga agactgcctc tatctgaata tctggtctcc caaacaggtc 420

ggtaaggggg ggctccccgt catgatctgg gtttacggcg gtgggtttag cggcggttct 480

ggcgcggtgc catattatga cggctctgcg ctcgcgcaga agggcgtggt ggtcgtcacg 540

ttcaactatc gcgccgggat tctgggcttt cttgcccatc cggcgctttc aaaggaaagt 600

ccgaatggcg tgtcgggcaa ctatggtctt ctcgacatgc tcgcggcgtt caaatgggtt 660

cagaacaaca taagggagtt cggcggagac ccgaaccgtg tcacggtctt tggcgagtcc 720

gccggcgcga gcgcgctcgg actgctcctg acctcgccgc tcagtgagag cgccttcaat 780

caggcgatac tgcaaagtcc gggtctggcc aggccgctcg ccacgctttc tgaaagcgaa 840

gcgaatgggc tggagctggg agccgatatt tctgctctac ggcgtgccga tgcgggcgaa 900

ttgacgaaga tcgcgcaatc gcgaataccc atgtcgcgcc agttcaccaa gccgcggccg 960

atgggtccga ttctggacgg ctatgttttg cgcacccttg acgtcgatgc cttcgccaag 1020

ggggccttcc gcaagatacc cgttctggtc ggcggaaacg ccgacgaagg gcgcgctttt 1080

acggatcgcc tgccggtcaa aacggtcctt gaatatcgag cctatctcac agaacaattt 1140

ggtgacgagg cggacgcatg ggagcgttgt tatcccgcga actccgacgc cgacgtcccc 1200

gccgccgttg cccgtctttt tggggatagt cagttcaaca acgggatcga gctgctctcg 1260

gcagccttcg cgaaatggcg aacgccgctt tggagatatc gctttacggg cattccagga 1320

gccggccgtc gccccgccac gcatggagac gaaattccct atgtcttcgc aaatctgggg 1380

ccgtcgtccg tatctatgtt tgggtcgctc gaaggcggcg ccggggcgtc ggacatcaaa 1440

cttgcgaccg aaatgtccgc ggcctgggtg agcttcgcgg tgcacggggt ccccgatcag 1500

ggcacgaaat cgcactggcc gcgcttcgag cggcgagggg agatcatgac ttttggttcg 1560

caggttggct ctggggaagg tcttggagtt tcgccgagca aagcctgcca accctcaaaa 1620

tag 1623

<210> 10

<211> 1503

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 10

ttggagtttc gccgagcaaa gcctgccaac cctcaaaata gcgcccggcc tgtgcgtgct 60

tcagcacgcc gtcccgcttt gcgggcgacg ggctgtgccc tctgcctaga aggaagtaag 120

ttgcgctacg acgtcgcgat aattggaggt ggcaacgctg cattgacggc agccgtgacg 180

gcgcgtgaag cgggggcctc ggttcttgtg atcgagcatg cgccgcgcgc catgcgcggc 240

ggcaacagtc gtcacacacg caatatgcgt acgatgcacg aacgtcccct gtcgccgttg 300

accggtgaat attcggcgga cgaatattgg aatgatcttg tccgcgtcac gggggggcgc 360

accgacgaag aactcgcgcg gctcgttatc cgcaacacca ccgacgctat tcccttcatg 420

acgcgctgcg gtgtgcgttt ccagccctcg ctgtcgggca cgctgagttt atcgcgaacc 480

aacgcattct tccttggcgg cgggaaggcg cttgtaaacg catattacgc cacggccgaa 540

cggctaggcg tcgatattct ctatgattct gaggtgaccg agatcaacct tcagcaaggc 600

gtcgtgcagc gtctgcaatt gcgcagccgg ggattccctg tcgaagtgga agccaaggct 660

gccatcgcct cgtccggagg attccaggca aatcttgact ggctctcaag cgcatggggg 720

cctgctgcgg cgaacttcat cgtacggggc acgccatatg cgactggcac ggtgctcaag 780

aacctgttgg agcaaggcgt cgcctcggtg ggagatccaa cccaatgcca tgctgtcgcg 840

atcgatgggc gagcgcccaa atacgacggc ggcatcgtca cacgactgga ctgcgttccc 900

ttctcgatcg tcgtcaacaa ggacgccttg cgcttctacg atgaaggcga agatgtgtgg 960

ccgaagcgtt acgccatatg gggtcgcttg gtggcacagc agcctgatca gatcgctttc 1020

agcataatcg atcggcaggc cgaagacctc ttcatgccgt cagtgttccc ccccgtgcaa 1080

gcggacacga tcgcgggtct ggccgagaaa ctcggtctga atcccgtaac cctggaacgc 1140

acggtggccg aattcaacgc cgcatgcgtg cccggcgaat tcggcggcca agatctcgac 1200

gacctccaca ccgagggaat cgaaccaaag aaatccaact gggcccgacc gattattgtg 1260

cccccgttca gcgcctatcc tctccggccc gggatcacct tcacctatct cggcgtcaag 1320

gtagacagcc gtgcgcgggt catcatggag acaggtgagc cgacaaaaaa cctgtttgct 1380

tcgggggaaa taatggcggg cagcattctc ggccaaggtt atctcgctgg atttggaatg 1440

gcgattggta ccgtattcgg ccgcatcgcg ggttgggagg ccgcacgtca tgcaggattt 1500

tga 1503

<210> 11

<211> 1173

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 11

atgcaggatt ttgatctcgt aaaaatgctg tctgacttgc cgtcggcgcc ggagctggaa 60

gccaggcgcg ttatggaggt gtgcaacgcg tgccgctatt gcgaagggtt ctgcgcggta 120

tttcctgcaa tgaccttgca gcgtcatttc gccagcggcg atctcagcca cctcgccaat 180

ctctgccact cgtgccaagg ttgctattac gcctgccaat acgcccctcc gcatgagttc 240

ggaataaacg ttccaaaggc gctgtcggag ttgcggctcg agagctacga gcagcatgct 300

tggccccggc cggtcgccgc tctctatcgc aagaatgcgc tcatcatttc catcttgtcg 360

gcggcatgca taaccggcgt ccttctgctt gccgccatct tcaacgggga tgcacttttc 420

gcgaaacacg catcggtgcc cggcggcggg ttttacaacg ttattcctta tcaggcgatg 480

attgccgtcg cggcgaccac atttctttat tccgcgctgg cgctggcgat cagtctcgtt 540

cgcttttcgc ggacgatcgg tctgggaatt aaggttcttt atcagcacgt gccggttctt 600

cgggcgctac gcgatgcggc gactctgcga tatctcggcg gcagcgacgg cgaggggtgt 660

aacgacgcgg acgagacatt ttcgacgacc cggcgaaaat ttcatcacgc ccttgcctat 720

ggcttcggac tttgtttcgc ggccacagcc acgggcacga tctacgatca tatgttcggc 780

tggccggcgc cctatgcgct tttcagcttg ccggtcgtcc tagggaccgt tggggggatc 840

ggaatggtcg tgggcgcgat cggcctactc tggctcaagc tggccggcga agacgctcct 900

cgatcaccgg cactgcttgg gccggatgtt gccctgttgg tgcttctgct tgccatagcg 960

gcaacgggcc tcctcctttt agcggtccgc agcaccgaag tcatgggcgt cgcgctcgcc 1020

gtccatctcg gcgtcgtctt ggccttcttt ttggtgatgc catacagcaa atttgtccac 1080

ggtatcttca ggctcacggc tctcgtgcgc catcatgctg accgcgaggc aagtaatggc 1140

ttcgcctcca gccctcccac gaaaaagggt taa 1173

<210> 12

<211> 1296

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 12

atggaacata tgaagtccgt tcgcgatcgc agtagcgtca tgcagatcgt gagagtggcg 60

agtggcaact gtctcgagca atatgatttc ttcgtttacg gcttctatgc ggcatatatt 120

gcgagaagct tttttccgac cggcgataac gcgacatcgc tcatgctttc attggccact 180

tttggcgctg gtttcctcat gaggcccttg ggggcgattt ttctcgggtc ctacatcgat 240

cgcgtcgggc gtcggaaagg cctgatcgtg acactcgcga tcatggccgt cggaaccctc 300

accattgcga tgactccaag ctatgaggca attggattac tcgcaccggt tatcgtgctc 360

gtcgggcgac ttttgcaggg tttttccgct ggagcagagt cgggtggcgt ctcagtgtac 420

ttggcggaaa ttgcgtcgcc caaatcgaga ggcttcttca cctcgtggca gtctgccagc 480

cagcaggtgg ccgtcatgat cgccgccgcg atcggtcttg cgctgcaatc aacgctttca 540

ccggagcaaa tgaacgactg gggatggcgg gtgcccttgt tgatcggatg cttgattatc 600

cccgtgatac tctggctgcg ccggtctctc ccggaaacga aagcctatct ccacatggag 660

cacaaggcgc attcgatcgg cgaatccctc cgcgaattgc aacagagctg ggggctgatc 720

ttgacgggca tggcgatgtc gatcctcacg acgaccacct tttacatgat taccgcctat 780

acgccgacat ttggcgagaa agcactcgga ctgagcccgc aagatgtcct gctggttacc 840

atcatggtcg gcgtgtcgaa cttcctgtgg cttccgatcg ggggtgctct ctcggatcgt 900

atcggtagaa ccccgatcct actggtcgtg ccggtcaccg ttctcgccat cgcctttccc 960

ctgatgagct ggctcgtcgc ggcaccgaca ttcggagcgc ttgcagctgt tctgctgact 1020

ttctccgcat gctttggact ctataatggg gcgctcatcg cgagactcac cgagattatg 1080

cctcccgcca ttagaaccct tggcttctcg ctggcgttca gtctcgcgac ctcgctgttc 1140

ggcggcttca ccccattggt aagtacggcg ctaatccacg cgacgggcag caattccgcg 1200

cctgcaatct ggctctgttt tgcggctttc atcagcttcg tcggtgtggc cgcatcgacc 1260

cggctgagcc ggccaatcgc cgaaggcgcc agatag 1296

<210> 13

<211> 1071

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 13

atgagagcag tagtttaccg aaatggcgaa cttgtcctgg gggcctatgc tgatccgata 60

cccgccgccg ggcaggtgct cgtcaagacc agagcatgcg gcatctgcgg atctgacctt 120

catttttgcg atcatgcgca ggcgtttacg aaccttgcat cgcgggcggg tatcgcctct 180

atggaagttg atttgtgtcg agacatcgtt ctggggcatg aattctgtgg cgagattatg 240

gagttcgggc cctctgcgga tcgtcgcttc aaacccggac agcttgtgtg ctcgctgccg 300

ctggcgatcg gtccgaccgg agcgcggacg attggctact cggatgagta tcccggcggg 360

ctcggcgaat atatggtcct cacggaagcg ctcttgctgc ctgttccgaa cggccttccg 420

gcgacctgcg cggcgttgac ggagccgatg gcggtgggat ggcatgccgt cgagatcgcg 480

caggttcaac cacatcacat ccctgtggtg atcgggtgcg gaccggtcgg gttggcagtc 540

gtcgctgccc tgaaacataa gcaagttgct ccgattattg cgtcggatcc atcgcccgat 600

cggcgtgctc ttgctctgcg gatgggcgcc gacgccgttg tcgatccgcg cgaagaatca 660

ccctttcgcc aggccgagaa gatcgcacgc ccggtcggac aaggtggggc cctgtccagc 720

tcattgctgt caaagtctca aatgatattc gaatgcgtag gggtgccggg catgcttcgg 780

catgcgatgg acggcgcgtc cgacgggtcc gagatcatgg tcgttggcgc atgcatgcag 840

ccggacgcga tcgagcccat gatcgggatg tttaaagcgc tcacgatcaa attctcgcga 900

acttacacgg gtgaggaatt cgccgcggtg cttcacatga taggtgaggg cgcactcgac 960

gtatctccgc tcgttaccga tgtgattggc ctgtccgatg tcccgtccgc gtttgaggct 1020

ctacggagtc caggcgccca agcaaaagtg attgtggacc cttggcgctg a 1071

<210> 14

<211> 1269

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 14

atggcgaacg gaacaaggca gaaagatctc agagaacgcg ccgaacgggt cattccgggc 60

gggatgtacg gccacgagtc gacacggttg ctgccgccag aattccccca gttcttcagg 120

cgcgcgctgg gggcacgaat ttgggacgcc gacgagcagc cctatatcga ctatatgtgc 180

gcgtatgggc caaatttgct cggttaccgg caatccgaaa tcgaagccgc ggctgatgcg 240

cagcgacttc tcggcgacac catgaccggt ccttcggaga tcatggtcaa cctcgccgaa 300

gcctttgtgg gcatggtccg tcatgcggat tgggcgatgt tctgcaaaaa tggcagcgat 360

gccacctcaa cggcgatggt tctcgcgcgt gcccatacgg ggcgcaaaac catattatgc 420

gccaaaggcg cctatcatgg cgcttccccg tggaacactc cgcatactgc cgggattctc 480

gcttccgatc gcgtgcatgt cgcatattat acctataacg acgcccaaag cttatcggac 540

gcgttcaagg cgcacgatgg cgatattgcg gctgtctttg ccacaccttt ccgacacgaa 600

gtatttgagg accaggccct cgcccagctt gagttcgcgc gcaccgctcg aaaatgttgt 660

gacgagaccg gtgcgcttct ggtcgttgac gatgtgcgcg caggtttccg ggtggcgcgc 720

gattgcagct ggacgcattt gggtatcgaa cccgatctca gttgctgggg aaaatgcttt 780

gcgaatggct atccgatctc cgccctgctg ggctcgaaca aggcgcgcga tgcggcgcgg 840

gatatatttg tgaccggctc cttctggttc tctgcggtac cgatggcggc cgcgatcgaa 900

accctcagga tcattcgaga gacgccttat ctcgaaacgc tgatcgccag cggcgccgcc 960

ctgcgggcag gcctggaggc acagtctcag cgccatggtc ttgagttgaa gcagacgggc 1020

ccggcgcaga tgccgcaaat attctttgcg gacgatcccg attttcggat cggctatgcg 1080

tgggccgcgg cgtgcctgaa gggcggcgtc tatgttcatc cctatcacaa tatgtttctc 1140

tctgcggccc atacagttga cgatgtaacg gagaccctcg aggcgacgga tcgcgcgttc 1200

agcgcggtcc tcagagattt tgcgtctctc cagcctcatc ccattttaat gcaactcgcc 1260

ggtgcttga 1269

<210> 15

<211> 2835

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 15

atgtatcgga agttcagaat cgaaaagccc ggcaaggcaa atagtttgct cggcgcagta 60

gcgctcggca ccctcgcatt tcctgtctct gccagtgctc aggatagcga tcccgcatcg 120

ataggtcagc cggacgaagc ggacacggac cggggaacga gcgaaatcgt cgtgaccggc 180

agccgcctcc agaacggctt caattcgccg acgccggtta cagccgtatc cagcgagcag 240

ttgaaggagg catctccgac caaccttgcc gacgcactca accagctgcc cgtgttcaac 300

gacagcttga agacctccaa ccctggcacg acacccggaa cggggaacag cggtcagaac 360

ctgctcaaca tgcgcggcct cgggtcaaac cggaacctcg tcctgctgaa cggcaaccgt 420

ttcgtcgcga ccaatttcac aggctcggtc gatatcaacg tgctgccgca ggcgttggtc 480

aagcgcgtcg atgtcgtgac gggcggcgcc tcggccgcct acggttccga tgccgtttcg 540

ggcgtcatca acttcgtgct cgacgaagat ctggaaggca tcagggccga gctccagtcg 600

ggtgtttcaa cccgcggcga cctcccgtcc tacggcggtt cgatcgcctt cggcacttcg 660

tttgccgacg accggttgca cttgctcggc agcttcgaat attttcgaca ggacggaatc 720

cgggccgatg aagcaacggg tcgccgctgg ttcgacatcg ccgccggcca atatcccgtg 780

cccggcgcta cgacaggcgt cacggtcgtg cccgatattc gcagttctcg cggatcctac 840

ggcggacttg tcacgtccgg ccctctgaaa ggcatcgcgt ttttgcccgg aggagtccta 900

gggaccttcg actacgggaa ttttacgagc tcgtcgttcc agagcggcgg cgatggaccg 960

cgcgtgaata tcggcttcgc cccggatcag cttcgctaca acgcgttcct acgcgccgca 1020

tatgatgtgt ccgacactgt gcaggtgtat gcggagggca cctatgctta ttcccacacc 1080

aacctgggtg cattcgtaat atcgcatgtc ggtggctcga ataatttccg gatcttccgt 1140

gataacgcct tccttccggc tccactcgcg acgctcatgg acagaaatgc ccaggcttcg 1200

atcgttgtcg gtcgcttctc aagcgacttt cccttggtcg aaatcgagaa tttcgcaaag 1260

gtctaccgcg gcgctgccgg cttccgggca gacattggca atggctggaa actcgatggc 1320

tcggcctcct ttggccttac ggacctcgag cttcgtgaaa acaatctcac catcaaccgc 1380

aatctctacg ccgccgtcga tgcggtccgc gatcccgcgg gcaatatcgt ctgccgttca 1440

acactggccg gcctcgacca agattgcgtg ccgctcaatc tcttcggcac aggctcgccg 1500

agcgcgtcgg ccatcgacta tgtcaccgct gatggcgtcg ctcagctgag gcttgagcaa 1560

tatgtggcgg gactcacgat ttccggcgac ctcggcgata gcctgtcgtt cggcgcgggc 1620

ccggtctcgg tcgccgctgg tatcgaatat cgcaaggaga aggcccggca ggaaaccgac 1680

gcgatatcgc aggcgacgac ctcgatcacg ggaatcaggg gggctccggc ggcgcaggca 1740

ggtcggcctg gaggcttcaa tctctacaac ccacttccct tctcgggaag ctatgacatc 1800

aaggaaggtt ttgtcgaaat cggcgtcccg attctgaagg acagcgcgct gggacgttcg 1860

ctgaacttaa acggcgccgt ccgatatgcc gattacagcc agtccggtgg agtaacaacc 1920

tggaagctgg gcggagaata tgagccgatc gacggcctca ggttccgcgc gacccgttcg 1980

cgagatatcc gcgggccaag ccttgtcgag ctattcgacc ccggccgtca ggcgacgctc 2040

aattcaattt atggcggaca ggctgtgcag acgcggttct ttaccgccgg caacgcggat 2100

ttgcgccctg aaaaggcgga cgtccttaca ttcggcgcgg tgctacgccc cgccttcgtg 2160

ccggggtttc agttttcggt cgatcgctat gtggtgaagg tgaagggcgc gatcgatttc 2220

ctccttcccc agcaggaaat cgacgcgtgc gatgcaggaa acaccttctt ctgcgacctc 2280

ataacggaga atccggacgg caccatcaca gtgacgggtc ccaatctcaa cctggctgtc 2340

cagaaagcgg cgggaattga cttcgaggcc tattactcac gccccgtcgg cggcggcacg 2400

ttcagtcttc gtgcgctggc aacgcaccat acctctgcct atcgcatcgc gaccggctcg 2460

gcgcccatcc gttcgctcgg acaaccggac acgccaaaat ggtcggccaa cttccaggcg 2520

cgatattcga ccgacgattg ggcgcttctc gtgcagcagc gcttcatcgc agcatcggtg 2580

ttcaatgccg acaatgtgga gggcgtcgat acgaatttga accacgctcc ggcggtttgg 2640

tacaccgacg cgacattgac cttcgacatc gcggcttttg gccagaagca gcagctgttt 2700

ctatcggtca ataatttgtt cgaccgagat ccgccaatag cgacgaacga ccccagcagt 2760

ttttccagcc cgaccagctc tgcctatgat ccggtcggcc gctattttaa tgtcggggtc 2820

cgtttccgga tctga 2835

<210> 16

<211> 551

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 16

atgcgcctca cgggcggaga attattggca cgatgtttgg ccgtcgaagg cgtccggtat 60

gtcttcggcc tcatgtcgcc ggaggtggat ccgctcctgg ctgcgctcga agacaatggg 120

atattgttcg tcccggtgcg gcacgaggcc gccgcagcct atatggccga gggcatttac 180

aagaccaccg gacaggtcgc cgcgattgtc acgaatccgg gtcccggtac ggcaaacctt 240

ctgcctggag tcgtgacggc acgccacgaa ggggttccct tcgtcgcaat aacgtcccag 300

catcaacttg gtgtcgttta tccctgcacg ccaaaaacct ttcagggaca agaccagatc 360

gacctctttc gacccgcggt taaatggggc gcacccatct tcgcctggaa ccggattgtc 420

gaaatcaccc atatggcgtt ccgggaaatg tgggccggca ggccgggacc cgttcagttg 480

gaaatcccga rgtctgtgat gtatgktgtg ggcgaacgag gaccacggta gaagtttacr 540

gatcgccgac a 551

<210> 17

<211> 1272

<212> DNA

<213> Artificial

<220>

<223> The sequence is synthesized.

<400> 17

atggaattga gccgccaacg agaccaggcc ttgagggagc gcgcccaagc ggtgatcccg 60

ggcgggatgt acggtcacga gtcgacctat ctgatgcccg agggcacgcc acagttcttc 120

agtcgcggca aaggcgcccg actttgggac gccgacggca acgagtatgt cgattacatg 180

tgcgcctatg gccccaacct gctgggttac ggcttcgaac ccgtcgaagc ggccgccgca 240

gcccagcaag cccggggcga taccctgacc gggccgtcgg aggtgatggt gcagttggcg 300

gaagacttcg tcgcgcaaat cagccacgcg gactgggcca tgttctgcaa gaacggcaca 360

gacgccacct caatggcgat ggtcatcgcg cgcgcacaca ccggccggaa gacgatcctc 420

tgcgcgaaag gcgcctatca tggggccgcg ccttggtgca cgccgatcct ggccggaacg 480

ctaccggagg atcgcgcctt tgtagtctac tacgactaca atgacgccca aagcctcgtc 540

gacgccttcg aggcccatca ggacgacgtc gcggcgatct tcgccacccc tcaccgtcac 600

gaggtgttca gcgaccagat cgatcctgat ccggaatatg cggccagcgt gcgggcgctc 660

tgcgacaaga gcggcgccct gctcgtcgtc gacgaagttc gagccgggtt caggatcgcg 720

cgcgactgca gctgggccaa gatcggcgtc gctccggatc tgagcacctg gggcaagtgc 780

ttcgccaacg gctatccgat ctcggcggtc ctagggggcg aaaaggtgcg cagcgcggca 840

aaggccgtct acgtcaccgg ctcgttctgg ttctcggcca cgcccatggc cgcagccgtc 900

gaaaccctga agcaaatccg cgagaccgac tatctcgagc ggatcaacgc ggccgggacc 960

cgcctgcgcg agggcctgca gcagcaggct gctcacaacg gctttacgtt gcgccaaacg 1020

gggcccgtct ccatgcccca agtcctcttc gaggaagatc ccgattttcg ggtcggctac 1080

ggctgggttc gcgaatgcct gaagcgaggg gtgtacttca gcccctacca taacatgttc 1140

ctgtcggcgg cccatagcga ggcggacctg gccaagaccc ttgcggctac cggcgacgcc 1200

ttcgtcgagc tacgcgccaa gcttccgagc ctagaaatcc accaacccct cctcgccctg 1260

agagcggcct aa 1272

Claims (10)

1. a kind of complex enzyme, which is characterized in that the complex enzyme contains amidase and esterase, which can remove fungi poison Element, particularly fumonisin, ochratoxin and T2 toxin.

2. complex enzyme according to claim 1, wherein, the amidase has：

(a)SEQ ID NO：Amino acid sequence shown in 2；

(b)SEQ ID NO：One or several in amino acid sequence shown in 2 in the 11st, 89,179 and 239 amino acids residues It is a that still there is SEQ ID NO after replacing, lacking or add：The amino acid sequence of amino acid sequence activity shown in 2；It is preferred that With by SEQ ID NOs：3rd, the nucleotide sequence coded amino acid sequence shown in 4 and 5.

3. complex enzyme according to claim 1, wherein, the esterase have SEQ ID NOs: 6,7,8,9,10,11, 12nd, the amino acid sequence encoded described in nucleotide sequence shown at least one of 13,14,15,16 and 17.

4. complex enzyme according to claim 1, wherein, the weight ratio of the amidase and esterase is 1：0.01-100.

5. complex enzyme according to claim 1, wherein, the complex enzyme also contains phosphatidylserine and/or Zn-ef ficiency.

6. complex enzyme according to claim 1, wherein, the complex enzyme also contains one or more other enzymes；It is described One or more other enzymes are selected from：Aflatoxin detoxifying enzymes, zearalenone lactonase, fumonisin Carboxylesterase, It is fumonisin aminopherase, amino polyol amine oxidase, deoxynivalenol epoxide hydrolase, carboxypeptidase, black Aspergillus aspartic protease PEPAa, PEPAb, PEPAc and PEPAd, elastoser, aminopeptidase, pepsin or stomach egg White enzyme sample protease, trypsase or trypsin like proteases, bacterialprotease are related to starch metabolism, fiber degradation, lipid The enzyme of metabolism, the protein for being related to glycogen metabolism or enzyme, amylase, arabinase, arabinofuranosidase, hydrogen peroxide Enzyme, cellulase, chitinase, renin, cutinase, deoxyribonuclease, epimerase, esterase, galactosidase, Portugal Dextranase, glucan lyase, endoglucanase, glucoamylase, glucose oxidase ,-glucosidase, including β-glucose Glycosides enzyme, glycuronidase, hemicellulase, hexoxidase, hydrolase, invertase, isomerase, lipolytic enzyme, laccase, cracking Enzyme, mannosidase, oxidizing ferment, oxidoreducing enzyme, pectate lyase, pectin acetyl esterases, pectin remove polymerase, pectinesterase Enzyme, pectin decomposing enzyme, peroxidase, phenol oxidase, phytase, polygalacturonase, protease, sandlwood-galactolipin Aldehydic acid enzyme, ribalgilase, African hesperidium element, transferase, transport protein, Transglutaminases, zytase, hexoxidase, Acid phosphatase and combination thereof.

7. a kind of additive, which is characterized in that the additive contains the complex enzyme described in any one in claim 1-6；

Preferably, on the basis of the total weight of additive, the dosage of the complex enzyme is 1-15ppm.

8. additive according to claim 7, wherein, the additive is also containing bacillus licheniformis, bacillus subtilis Bacterium, bifidobacterium bifidum, enterococcus faecalis, enterococcus faecium, lactoenterococcus, lactobacillus acidophilus, Lactobacillus casei, German-style lactobacillus Lactic acid subspecies, lactobacillus plantarum, Pediococcus acidilactici, Pediococcus pentosaceus, candida utili, bifidobacterium infantis, long bifid bar Bacterium, bifidobacterium breve, bifidobacterium adolescentis, streptococcus thermophilus, lactobacillus reuteri, animal bifidobacteria, aspergillus oryzae, slow bud In spore bacillus, bacillus pumilus, lactobacillus cellobiosas, lactobacillus fermenti and lactobacillus delbruockii subspecies bulgaricus at least It is a kind of；

Preferably, the additive is used for ensilage or ox feed, and the additive is also containing production propionibacterium acide-propionici, Bu Shi At least one of lactobacillus and Lactobacillus paracasei；Or

The additive is used for the feed of poultry, growing-finishing pig and aquiculture animal, and the additive is also containing condensation bud Spore bacillus and/or Brevibacillus laterosporus.

9. the complex enzyme described in any one and/or the additive described in claim 7 or 8 are true in removing in claim 1-6 Application in verticillium toxin particularly fumonisin, ochratoxin and T2 toxin.

A kind of 10. method for removing mycotoxin, which is characterized in that this method includes：By any one institute in claim 1-6 The additive described in complex enzyme and/or claim 7 or 8 stated is contacted with the sample that mycotoxin pollutes, to remove in sample Mycotoxin, the mycotoxin is preferably fumonisin, ochratoxin and T2 toxin.