WO2022007145A1 - Gene module for efficiently degrading feather and synthesizing artificial hemeprotein, and application thereof - Google Patents

Gene module for efficiently degrading feather and synthesizing artificial hemeprotein, and application thereof Download PDF

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WO2022007145A1
WO2022007145A1 PCT/CN2020/111685 CN2020111685W WO2022007145A1 WO 2022007145 A1 WO2022007145 A1 WO 2022007145A1 CN 2020111685 W CN2020111685 W CN 2020111685W WO 2022007145 A1 WO2022007145 A1 WO 2022007145A1
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林敏�
周正富
燕永亮
张维
王劲
陆伟
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中国农业科学院生物技术研究所
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  • the invention belongs to the technical field of synthetic biology biotransformation, and relates to a high-efficiency artificially designed biosynthesis system for degrading animal wastes such as feathers and converting synthetic heme protein.
  • Livestock wastes such as feathers, furs, hooves, horns, etc.
  • the massive accumulation of these wastes not only becomes a potential source of environmental pollution, but also causes a huge waste of resources.
  • these wastes are rich in protein and amino acids and are potential protein and energy resources.
  • Using modern synthetic biology protein intelligent design methods to create an efficient biotransformation cell factory for animal feather organic waste can accelerate the solution to the problem of massive accumulation of livestock waste, and realize waste reduction and efficient utilization of resources.
  • Synthetic biology is based on the intersection of biology, chemistry, physics, computing, engineering and other disciplines. It redesigns or even de novo synthesis of organisms in an engineered way, which will overcome the limitations of natural evolution and create synthetic organisms that surpass natural life capabilities.
  • the purpose of the present invention is to use synthetic biology methods to create an efficient biosynthetic system for degrading animal waste such as feathers and converting them into heme protein.
  • the invention creates an efficient biosynthesis system for converting animal waste into artificial heme protein, and provides a functional module capable of degrading poultry feather waste and synthesizing heme protein, which is named SyKerMb.
  • the SyKerMb module is the nucleotide of the sequence shown in SEQ ID NO.3.
  • the size of the full-length nucleic acid sequence of SEQ ID NO.3 is 1635bp, and this module encodes 544 amino acids.
  • SEQ ID NO.3 is assembled by artificially designed nucleotides of the sequences shown in SEQ ID NO.1 and SEQ ID NO.2.
  • SEQ ID NO.1 is the sequence designed and synthesized for the first time in the present invention.
  • nucleic acid sequence of functional module SyKerMb was obtained by artificial chemical synthesis. Its size is 1635bp nucleotides, and the modular product consists of 544 amino acids;
  • SyKerMb was cloned on the vector pJET, and the recombinant clone plasmid pJET-SyKerMb containing the complete functional module SyKerMb was constructed;
  • the biosynthetic functional module SyKerMb is connected to the expression vector pET28t plasmid, the plasmid contains the inducible T7 promoter, the target module protein that can be expressed in the presence of the inducer IPTG.
  • the heme protein obtained above was shown by polyacrylamide gel electrophoresis (SDS-PAGE), and after purification, the protein molecular weight was about 18Kd.
  • SEQ ID NO.1 artificially designed new nucleotide sequence, assembling one of the nucleotides of SEQ ID NO.3;
  • SEQ ID NO.2 another nucleotide that assembles SEQ ID NO.3;
  • SEQ ID NO.3 the nucleotide sequence of the functional module SyKerMb;
  • SEQ ID NO. 4 Amino acid sequence of the protein encoded by SyKerMb.
  • Fig. 1 Degradation and transformation effect of engineering strain BL21-SyKerMb containing module SyKerMb on feather substrate;
  • FIG. 1 SDS-PAGE gel image of the engineered strain BL21-SyKerMb containing the module SyKerMb. 1. The supernatant of the whole protein fragmentation liquid of the strain induced for 24h; 2, the supernatant of the whole protein fragmentation liquid of the strain induced for 96h; 3, the whole protein of the control strain.
  • Plasmids, bacterial strain sources cited in the embodiment are as follows:
  • Cloning vector pJET a commercially available product from ThermoFisher;
  • Escherichia coli BL21 (DE3): a commercial product of Beijing Quanshijin Company.
  • Feather Procured from the market for chicken feathers.
  • IPTG isopropyl-beta-D-thiogalactoside.
  • Cloning vector pJET a commercially available product from ThermoFisher;
  • Escherichia coli BL21 (DE3): a commercial product of Beijing Quanshijin Company.
  • the full-length nucleic acid sequence of the functional module SyKerMb was obtained by means of artificial chemical synthesis through the synthetic biology design module. Its size is 1635bp, and the module consists of 544 amino acids. It was cloned into the vector pJET, and the recombinant clone plasmid pJET-SyKerMb containing the complete functional module SyKerMb was constructed and verified by sequencing; The functional module SyKerMb containing sticky ends and the expression vector pET-28t vector containing the T7 promoter were obtained, the functional module SyKerMb was connected to the pET-28t vector, the high expression vector pET28t-SyKerMb of E. coli was constructed, and the expression vector was transformed into E. coli BL21(DE3).
  • the full-length nucleic acid sequence of the functional module SyKerMb was obtained by artificial chemical synthesis, and a recombinant E. coli engineering strain expressing the functional module SyKerMb was successfully constructed. After PCR, enzyme digestion, and sequencing, the inserted sequence was verified to be correct, and the strain was named BL21-SyKerMb. E. coli BL21 (DE3) containing the pET-28t control empty plasmid was named BL21-28.
  • Recombinant engineering strain the engineering strain BL21-SyKerMb strain expressing the SyKerMb functional module obtained in Example 1
  • Control strain the BL21-28 strain containing the empty plasmid described in Example 1.
  • Inorganic salt medium NaCl 0.05%, K 2 HPO 4 0.1%, KH 2 PO 4 0.04%, MgCl 2 ⁇ 7H 2 O 0.01%, CaCl 2 0.006%, pH 7.5.
  • the seed solution of the engineered strain expressing the module SyKerMb was transferred to an inorganic salt medium with feathers as the sole carbon and nitrogen source, and the inducer IPTG (final concentration 0.1 mM) was added. Feather degradation.
  • the engineered strain BL21-SyKerMb was cultured with feathers as the sole carbon and nitrogen source, and the inducer IPTG (final concentration 0.5 mM) was added.
  • the cells of the strain were lysed by an ultrasonic cell disruptor, and the supernatant and precipitate of the cell disruption liquid were collected by centrifugation. A small amount of samples were taken for SDS-PAGE electrophoresis to detect and identify the protein expression of the engineered strain BL21-SyKerMb.

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Abstract

A functional module SyKerMb capable of efficiently degrading poultry feather waste and synthesizing a novel hemeprotein is artificially designed and constructed by using a synthetic biology method. A recombinant vector of the functional module is constructed, and assembled and expressed in chassis microorganism cell Escherichia coli. Experiments prove that engineering strain integrating the transformation function module has the biological synthesis capability of degrading animal feather waste and efficiently synthesizing an artificial hemeprotein, and can be applied in related industrial production of artificial protein preparations and future artificial food.

Description

一种高效降解羽毛并合成人工血红素蛋白的基因模块及应用A Gene Module and Application for Efficiently Degrading Feather and Synthesizing Artificial Heme Protein 技术领域technical field
本发明属于合成生物学生物转化技术领域，涉及一种降解动物废弃物如羽毛，转化合成血红素蛋白的高效人工设计生物合成体系。The invention belongs to the technical field of synthetic biology biotransformation, and relates to a high-efficiency artificially designed biosynthesis system for degrading animal wastes such as feathers and converting synthetic heme protein.
背景技术Background technique
禽畜废弃物，如羽毛、毛皮、蹄、角等是过去长期无法利用的农业有机废弃物。这些废弃物大量累积，不但成为潜在的环境污染源，更造成了极大的资源浪费。然而，这些废弃物中蛋白质和氨基酸的含量丰富，是潜在的蛋白质和能量资源。利用现代合成生物学蛋白智能设计方法，创建动物羽毛有机废弃物的高效生物转化细胞工厂，能够加速解决禽畜废弃物大量堆积问题，实现废弃物减排与资源化的高效利用。Livestock wastes, such as feathers, furs, hooves, horns, etc., are agricultural organic wastes that could not be used for a long time in the past. The massive accumulation of these wastes not only becomes a potential source of environmental pollution, but also causes a huge waste of resources. However, these wastes are rich in protein and amino acids and are potential protein and energy resources. Using modern synthetic biology protein intelligent design methods to create an efficient biotransformation cell factory for animal feather organic waste can accelerate the solution to the problem of massive accumulation of livestock waste, and realize waste reduction and efficient utilization of resources.
合成生物学基于生物、化学、物理、计算、工程等多学科交叉，对生物体以工程化的方式重新设计甚至是从头合成，将克服自然进化的局限，创造超越自然生命能力的合成生物。Synthetic biology is based on the intersection of biology, chemistry, physics, computing, engineering and other disciplines. It redesigns or even de novo synthesis of organisms in an engineered way, which will overcome the limitations of natural evolution and create synthetic organisms that surpass natural life capabilities.
发明内容SUMMARY OF THE INVENTION
本发明的目的是利用合成生物学方法创建一种降解动物废弃物如羽毛，并将其转化为血红素蛋白的高效生物合成体系。The purpose of the present invention is to use synthetic biology methods to create an efficient biosynthetic system for degrading animal waste such as feathers and converting them into heme protein.
本发明创建了将动物废弃物转化为人工血红素蛋白的高效生物合成体系，提供了一种能够降解家禽羽毛废弃物并合成血红素蛋白的功能模块，命名为SyKerMb。The invention creates an efficient biosynthesis system for converting animal waste into artificial heme protein, and provides a functional module capable of degrading poultry feather waste and synthesizing heme protein, which is named SyKerMb.
所述SyKerMb模块是SEQ ID NO.3所示序列的核苷酸。SEQ ID NO.3全长核酸序列大小为1635bp，该模块编码544个氨基酸。The SyKerMb module is the nucleotide of the sequence shown in SEQ ID NO.3. The size of the full-length nucleic acid sequence of SEQ ID NO.3 is 1635bp, and this module encodes 544 amino acids.
所述SEQ ID NO.3由人工设计的SEQ ID NO.1和SEQ ID NO.2所示序列的核苷酸组装而成。Described SEQ ID NO.3 is assembled by artificially designed nucleotides of the sequences shown in SEQ ID NO.1 and SEQ ID NO.2.
其中，SEQ ID NO.1为本发明首次设计合成的序列。Wherein, SEQ ID NO.1 is the sequence designed and synthesized for the first time in the present invention.
具体研究工作如下：The specific research work is as follows:
1、获得功能模块SyKerMb全长核酸序列1. Obtain the full-length nucleic acid sequence of the functional module SyKerMb
通过合成生物学设计，利用人工化学合成的方法获得了功能模块SyKerMb全长核酸序列。其大小为1635bp核苷酸，该模块产物由544个氨基酸组成；Through synthetic biology design, the full-length nucleic acid sequence of functional module SyKerMb was obtained by artificial chemical synthesis. Its size is 1635bp nucleotides, and the modular product consists of 544 amino acids;
2、构建含有功能模块SyKerMb的重组工程菌株2. Construction of recombinant engineering strains containing functional module SyKerMb
1)将SyKerMb克隆于载体pJET上，构建了含有完整功能模块SyKerMb的重组克隆质粒pJET-SyKerMb；1) SyKerMb was cloned on the vector pJET, and the recombinant clone plasmid pJET-SyKerMb containing the complete functional module SyKerMb was constructed;
2)将生物合成功能模块SyKerMb连接于表达载体pET28t质粒上，该质粒含诱导型T7启动子，可在诱导物IPTG存在时表达的靶标模块蛋白。构建完成的重组质粒pET28t-SyKerMb；2) The biosynthetic functional module SyKerMb is connected to the expression vector pET28t plasmid, the plasmid contains the inducible T7 promoter, the target module protein that can be expressed in the presence of the inducer IPTG. The constructed recombinant plasmid pET28t-SyKerMb;
3)将导入功能模块SyKerMb的重组质粒pET28t-SyKerMb转入受体大肠杆菌BL21(DE3)中，获得高效生物合成工程菌株BL21-SyKerMb(详见实施例1)；3) The recombinant plasmid pET28t-SyKerMb introduced into the functional module SyKerMb was transferred into the recipient Escherichia coli BL21 (DE3) to obtain a high-efficiency biosynthetic engineering strain BL21-SyKerMb (see Example 1 for details);
3、模块SyKerMb的功能验证3. Functional verification of module SyKerMb
用上述含有模块SyKerMb的工程菌株进行了如下生物降解与生物合成实验：The following biodegradation and biosynthesis experiments were carried out with the above engineering strains containing the module SyKerMb:
1)废弃物羽毛降解转化与生物合成实验1) Degradation, transformation and biosynthesis experiments of waste feathers
实验结果显示，羽毛在含有模块SyKerMb的工程菌株培养液中，从24h起，开始大量分解；培养96h时菌株细胞呈现鲜红色，胞内大量积累血红素蛋白；The experimental results showed that feathers began to decompose in large quantities in the culture solution of the engineering strain containing the module SyKerMb from 24h; after culturing for 96h, the cells of the strain appeared bright red, and a large amount of heme protein was accumulated in the cells;
不含SyKerMb的对照菌株BL21-28培养液中，羽毛未见降解发生(详见实施例2)。In the culture medium of the control strain BL21-28 without SyKerMb, no feather degradation occurred (see Example 2 for details).
2)转化得到的人工蛋白的纯化与分子量测定2) Purification and molecular weight determination of artificial protein obtained by transformation
将前述获得的血红素蛋白经聚丙烯酰胺凝胶电泳(SDS-PAGE)显示，所述蛋白纯化后，蛋白分子量约18KdThe heme protein obtained above was shown by polyacrylamide gel electrophoresis (SDS-PAGE), and after purification, the protein molecular weight was about 18Kd.
说明，表达模块SyKerMb的工程菌株能有效降解羽毛废弃物，并转化合成人工血红素蛋白。It shows that the engineered strain expressing the module SyKerMb can effectively degrade feather waste and transform into synthetic artificial heme protein.
序列表信息Sequence Listing Information
SEQ ID NO.1：人工设计的新核苷酸序列，组装SEQ ID NO.3的核苷酸之一；SEQ ID NO.1: artificially designed new nucleotide sequence, assembling one of the nucleotides of SEQ ID NO.3;
SEQ ID NO.2：组装SEQ ID NO.3的另一核苷酸；SEQ ID NO.2: another nucleotide that assembles SEQ ID NO.3;
SEQ ID NO.3：功能模块SyKerMb的核苷酸序列；SEQ ID NO.3: the nucleotide sequence of the functional module SyKerMb;
SEQ ID NO.4：SyKerMb编码蛋白的氨基酸序列。SEQ ID NO. 4: Amino acid sequence of the protein encoded by SyKerMb.
附图说明：Description of drawings:
图1含模块SyKerMb的工程菌株BL21-SyKerMb对羽毛底物的降解转化效果；Fig. 1 Degradation and transformation effect of engineering strain BL21-SyKerMb containing module SyKerMb on feather substrate;
图2含模块SyKerMb的工程菌株BL21-SyKerMb的SDS-PAGE胶图。1，诱导 24h菌株全蛋白破碎液上清；2，诱导96h菌株全蛋白破碎液上清；3对照菌株全蛋白。Figure 2. SDS-PAGE gel image of the engineered strain BL21-SyKerMb containing the module SyKerMb. 1. The supernatant of the whole protein fragmentation liquid of the strain induced for 24h; 2, the supernatant of the whole protein fragmentation liquid of the strain induced for 96h; 3, the whole protein of the control strain.
具体实施方式detailed description
以下实施例中所举的质粒、菌株以及微生物催化降解的对象只用于对本发明作进一步详细说明，并不对本发明的实质内容加以限制。凡未注明具体实验条件的，均为按照本领域技术人员熟知的常规条件或按照制造厂商所建议的条件。实施例中所举的质粒、菌株来源如下：The plasmids, strains and the objects of catalyzed degradation by microorganisms in the following examples are only used to further describe the present invention in detail, and do not limit the essential content of the present invention. Where the specific experimental conditions are not indicated, all are in accordance with the conventional conditions well known to those skilled in the art or in accordance with the conditions suggested by the manufacturer. Plasmids, bacterial strain sources cited in the embodiment are as follows:
克隆载体pJET：为ThermoFisher公司市售产品；Cloning vector pJET: a commercially available product from ThermoFisher;
穿梭质粒pET-28t：本实验室保存；Shuttle plasmid pET-28t: kept in our laboratory;
大肠杆菌BL21(DE3)：为北京全式金公司市售产品。Escherichia coli BL21 (DE3): a commercial product of Beijing Quanshijin Company.
羽毛：为鸡羽毛采购于市场。Feather: Procured from the market for chicken feathers.
IPTG：异丙基-β-D-硫代半乳糖苷。IPTG: isopropyl-beta-D-thiogalactoside.
实施例1含有转化功能模块SyKerMb的重组工程菌株构建Example 1 Construction of recombinant engineering strains containing transformation functional module SyKerMb
一、实验材料1. Experimental materials
克隆载体pJET：为ThermoFisher公司市售产品；Cloning vector pJET: a commercially available product from ThermoFisher;
表达质粒pET-28t：本实验室保存；Expression plasmid pET-28t: kept in our laboratory;
大肠杆菌BL21(DE3)：为北京全式金公司市售产品。Escherichia coli BL21 (DE3): a commercial product of Beijing Quanshijin Company.
二、实验方法2. Experimental method
1.通过合成生物学设计模块，利用人工化学合成的方法获得了功能模块SyKerMb全长核酸序列。其大小为1635bp，该模块由544个氨基酸组成，将其克隆于载体pJET上，构建了含有完整功能模块SyKerMb的重组克隆质粒pJET-SyKerMb，并测序验证；然后通过Nde I/Xho I双酶切获得含有粘性末端的功能模块SyKerMb及含有T7启动子的表达载体pET-28t载体，将功能模块SyKerMb连接于pET-28t载体上，构建大肠杆菌高表达载体pET28t-SyKerMb，将该表达载体转化大肠杆菌BL21(DE3)。1. The full-length nucleic acid sequence of the functional module SyKerMb was obtained by means of artificial chemical synthesis through the synthetic biology design module. Its size is 1635bp, and the module consists of 544 amino acids. It was cloned into the vector pJET, and the recombinant clone plasmid pJET-SyKerMb containing the complete functional module SyKerMb was constructed and verified by sequencing; The functional module SyKerMb containing sticky ends and the expression vector pET-28t vector containing the T7 promoter were obtained, the functional module SyKerMb was connected to the pET-28t vector, the high expression vector pET28t-SyKerMb of E. coli was constructed, and the expression vector was transformed into E. coli BL21(DE3).
三、实验结果3. Experimental results
利用人工化学合成的方法获得了功能模块SyKerMb全长核酸序列，成功构建了表达功能模块SyKerMb的重组大肠杆菌工程菌株。经PCR、酶切，测序验证***序列正确，将该菌株命名为BL21-SyKerMb。含有pET-28t对照空质粒的E.coli BL21(DE3)命名为BL21-28。The full-length nucleic acid sequence of the functional module SyKerMb was obtained by artificial chemical synthesis, and a recombinant E. coli engineering strain expressing the functional module SyKerMb was successfully constructed. After PCR, enzyme digestion, and sequencing, the inserted sequence was verified to be correct, and the strain was named BL21-SyKerMb. E. coli BL21 (DE3) containing the pET-28t control empty plasmid was named BL21-28.
四、实验结论Fourth, the experimental conclusion
完成表达SyKerMb功能模块的重组大肠杆菌工程菌株的构建。The construction of recombinant E. coli engineering strain expressing SyKerMb functional module was completed.
实施例2含有模块SyKerMb高效生物合成工程菌株的生物转化检测Example 2 Biotransformation detection of high-efficiency biosynthetic engineered strains containing module SyKerMb
一、实验材料1. Experimental materials
重组工程菌株：实施例1得到的表达SyKerMb功能模块的工程菌株BL21-SyKerMb菌株Recombinant engineering strain: the engineering strain BL21-SyKerMb strain expressing the SyKerMb functional module obtained in Example 1
对照菌株：实施例1所述含空质粒的BL21-28菌株。Control strain: the BL21-28 strain containing the empty plasmid described in Example 1.
二、实验方法2. Experimental method
1.废弃物羽毛降解实验1. Waste feather degradation experiment
用清水冲洗鸡羽毛，烘干备用。以羽毛为唯一碳源和氮源加入50mL无机盐培养基，高温灭菌。无机盐培养基(m/v)：NaCl 0.05％，K 2HPO 4 0.1％，KH 2PO 4 0.04％，MgCl 2·7H 2O 0.01％，CaCl 2 0.006％，pH7.5。 Rinse chicken feathers with clean water and dry for later use. Using feathers as the sole carbon and nitrogen source, 50 mL of inorganic salt medium was added, and sterilized at high temperature. Inorganic salt medium (m/v): NaCl 0.05%, K 2 HPO 4 0.1%, KH 2 PO 4 0.04%, MgCl 2 ·7H 2 O 0.01%, CaCl 2 0.006%, pH 7.5.
将表达模块SyKerMb的工程菌株种子液转接于以羽毛为唯一碳源和氮源加入无机盐培养基中，加入诱导物IPTG(终浓度0.1mM)，37℃恒温摇床培养，每隔12h观测羽毛降解情况。The seed solution of the engineered strain expressing the module SyKerMb was transferred to an inorganic salt medium with feathers as the sole carbon and nitrogen source, and the inducer IPTG (final concentration 0.1 mM) was added. Feather degradation.
2.人工血红素蛋白的表达与纯化实验2. Expression and purification experiments of artificial heme protein
将以羽毛为唯一碳源和氮源培养工程菌株BL21-SyKerMb，加入诱导物IPTG(终浓度0.5mM)，37℃恒温摇床培养，每隔24h收集离心菌株。利用超声波细胞破碎仪裂解菌株细胞，离心分别收集细胞破碎液上清与沉淀，取少量样品进行SDS-PAGE电泳，检测和鉴定工程菌株BL21-SyKerMb的蛋白表达情况。The engineered strain BL21-SyKerMb was cultured with feathers as the sole carbon and nitrogen source, and the inducer IPTG (final concentration 0.5 mM) was added. The cells of the strain were lysed by an ultrasonic cell disruptor, and the supernatant and precipitate of the cell disruption liquid were collected by centrifugation. A small amount of samples were taken for SDS-PAGE electrophoresis to detect and identify the protein expression of the engineered strain BL21-SyKerMb.
三、实验结果3. Experimental results
羽毛降解实验显示，菌株BL21-SyKerMb培养24h已经开始大量降解羽毛，48h后菌株已经大量生长并呈现淡粉色，96h，菌株胞内大量积累人工血红素蛋白，菌株细胞呈现鲜红色(图1)。SDS-PAGE结果显示，培养重组工程菌株大量48h时，细胞内已经合成大量人工血红素蛋白；培养48h后，胞内肌浆蛋白大量积累。人工血红素蛋白分子量约18kD(图2)。研究结果显示，表达模块SyKerMb的工程菌株能够降解羽毛废弃物来提供碳源和氮源正常生长，同时胞内能够合成靶标人工血红素蛋白。Feather degradation experiments showed that the strain BL21-SyKerMb had begun to degrade feathers in a large amount after 24 hours of culture. After 48 hours, the strain had grown a lot and turned pale pink. At 96 hours, artificial heme protein was accumulated in the cells of the strain, and the cells of the strain appeared bright red (Figure 1). The results of SDS-PAGE showed that a large amount of artificial heme protein had been synthesized in the cells when the recombinant engineered strain was cultured for 48 hours; after 48 hours of culture, a large amount of intracellular sarcoplasmic protein was accumulated. The artificial heme protein has a molecular weight of about 18kD (Figure 2). The results show that the engineered strain expressing the module SyKerMb can degrade feather waste to provide carbon and nitrogen sources for normal growth, and at the same time, it can synthesize the target artificial heme protein intracellularly.
四、实验结论Fourth, the experimental conclusion
实验表明，模块SyKerMb的表达使工程菌株能够将羽毛废弃物转化合成靶标人工血红素蛋白。Experiments show that the expression of the module SyKerMb enables the engineered strain to convert feather waste into synthetic target artificial heme proteins.

Claims (9)

  1. SEQ ID NO:1所示核苷酸序列的DNA。DNA of the nucleotide sequence shown in SEQ ID NO: 1.
  2. 含SEQ ID NO:1所示核苷酸序列的DNA的质粒。A plasmid containing the DNA of the nucleotide sequence shown in SEQ ID NO: 1.
  3. 含SEQ ID NO:1所示核苷酸序列的DNA的重组工程菌。Recombinant engineering bacteria containing the DNA of the nucleotide sequence shown in SEQ ID NO: 1.
  4. 含有SEQ ID NO:1所示核苷酸序列的功能模块在催化生物降解转化合成中的应用。The application of the functional module containing the nucleotide sequence shown in SEQ ID NO: 1 in the synthesis of catalytic biodegradation conversion.
  5. 权利要求4所述的应用，所述的转化合成是将禽类羽毛降解并转化为人工血红素蛋白的生物合成反应。The application according to claim 4, wherein the transformation synthesis is a biosynthetic reaction of degrading and transforming avian feathers into artificial heme proteins.
  6. 一种降解羽毛并将其转化为血红素蛋白的功能模块，由SEQ ID NO:3所示序列的DNA组成。A functional module that degrades feathers and converts them into heme proteins, consisting of DNA with the sequence shown in SEQ ID NO:3.
  7. SEQ ID NO:3所示序列的DNA在催化生物降解转化合成中的应用。The application of the DNA of the sequence shown in SEQ ID NO: 3 in catalyzing biodegradation, transformation and synthesis.
  8. 权利要求7所述的应用，所述的转化合成是将禽类羽毛降解并转化为人工血红素蛋白的生物合成反应。The application according to claim 7, wherein the transformation synthesis is a biosynthetic reaction of degrading poultry feathers and converting them into artificial heme proteins.
  9. SEQ ID NO:3所示序列的功能模块编码的氨基酸序列，如SEQ ID NO:4所示。The amino acid sequence encoded by the functional module of the sequence shown in SEQ ID NO:3 is shown in SEQ ID NO:4.
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