895 results about "Bioengineerings" patented technology

The invention discloses an arginine decarboxylase diaA enzyme mutant and application thereof in preparation of butanediamine, and belongs to the field of bioengineering. According to the invention, rational charge overturning transformation is simultaneously carried out on a pentamer meridian oligomeric interface and a dimer latitudinal oligomeric interface, so that stable assembly and efficient catalysis of the decamer under the condition of neutral to alkaline pH (7.0-9.0) are realized. Wherein positive charges are introduced into a meridian interface to weaken electrostatic repulsion, and negative charges are introduced into a latitudinal interface to enhance dimer compactness and substrate transfer efficiency. The specific enzyme activity of the representative double mutant AdiAD110K / H736E at pH 8.0 is about 35 times that of a wild type, and the representative double mutant AdiAD110K / H736E keeps a complete decamer state in a pH range of 7.0-9.0. The yield of butanediamine is up to 145.9 g / L under the whole-cell catalysis of the mutant.

The invention discloses a rational copolymerization strategy for improving the stability of an oligomeric structure of acid-induced high-order oligomeric decarboxylase AdiA in a neutral to alkaline environment and application of the rational copolymerization strategy, and belongs to the field of bioengineering. According to the strategy, rational charge overturning transformation is carried out on a pentamer radial oligomeric interface and a dimer weft-wise oligomeric interface at the same time, and stable assembly and efficient catalysis of a decamer under the condition that the pH value is from 7.0 to 9.0 from neutral to alkaline are achieved. Wherein positive charges are introduced into a meridian interface to weaken electrostatic repulsion, and negative charges are introduced into a latitudinal interface to enhance dimer compactness and substrate transfer efficiency. The specific enzyme activity of the representative double mutant AdiAD471K / E467K / H736E is 45.5 times that of a wild type when the pH value is 8.0, and the representative double mutant AdiAD471K / E467K / H736E keeps a complete decamer state when the pH value is 7.0-9.0. The yield of butanediamine is up to 156.5 g / L by using the mutant to catalyze whole cells.

The invention relates to a CHO-S cell strain capable of stably expressing H5N1 hemagglutinin protein and a construction method of the CHO-S cell strain, and belongs to the field of bioengineering.H5N1 hemagglutinin protein expression plasmids are obtained in an in-vitro synthesis and seamless cloning mode, 293T cells are transfected through the H5N1 hemagglutinin protein expression plasmids, and it is proved that the H5N1 hemagglutinin protein is expressed; the CHO-S cell strain capable of stably expressing the H5N1 hemagglutinin protein is obtained by transfecting CHO-S cells by using the H5N1 hemagglutinin protein expression plasmids and carrying out multiple rounds of cloning and screening, so that a basis is provided for obtaining H5N1 recombinant protein influenza vaccines.

The invention discloses an aspergillus niger recombinant bacterium AnCT-XynB-K-agdA and application thereof, and belongs to the technical field of microbiology and bioengineering. According to the invention, Aspergillus niger AnCat is taken as an expression host, firstly, an acid protease regulatory factor prtT is knocked out to obtain a defective strain, and then different chaperonins are subjected to fusion expression by optimizing integration sites of a laccase lcc9 expression cassette, so that the high-expression laccase chaperonin is obtained. According to the method, an expression vector pC3-5 'agdA-PcitA-XynB-L-lcc9-hph-3' agdA is constructed, a recombinant bacterium AnCT-XynB-K-agdA is further obtained, the enzyme activity of the extracellular laccase subjected to shake flask fermentation reaches 1821.2 U / L, and finally, the recombinant efficient expression of the laccase Lcc9 in aspergillus niger is realized.

The invention belongs to the technical field of bioengineering, and particularly relates to leuconostoc mesenteroides subsp. Mesenteroides FTCM002 and application thereof in preparation of dendrobium officinale leavening with the effects of reducing blood sugar, nourishing the stomach and resisting inflammation. Dendrobium officinale is subjected to enzymolysis and inoculated with the strain for fermentation, and the obtained fermented dendrobium officinale has the effect of inhibiting alpha-glucosidase; the gastric mucosal injury can be relieved, the expression of VEGF, IL-6, IL-8 and TNF-alpha in the gastric mucosal injury is reduced, and the effect of nourishing the stomach is achieved; the cell phagocytic rate can be increased, the NO concentration and the content of immune factors IL-6 and TNF-alpha can be increased, and the effect of enhancing the immunity is achieved; the expression quantity of IL-6, IL-1beta and TNF-alpha in inflammatory cells can be reduced, and the anti-inflammatory effect is achieved.

The invention discloses a GmbZIP36 protein related to aluminum resistance of plants in acid soil as well as a coding gene and application of the GmbZIP36 protein, and belongs to the technical field of bioengineering. The amino acid sequence of the GmbZIP36 protein is shown as SEQ ID NO: 2, 3 or 4, and the nucleotide sequence of the coding gene of the GmbZIP36 protein is shown as SEQ ID NO: 1. The invention finds that the expression of the GmbZIP36 gene is induced by aluminum stress, the relative root elongation and root biomass of arabidopsis thaliana and soybean under the aluminum stress can be obviously improved by over-expressing the gene in a plant, and aluminum accumulation is reduced, so that the aluminum resistance of the plant is enhanced. The invention provides a recombinant vector containing the gene, a host cell and application of the recombinant vector and the host cell in regulating and controlling the aluminum resistance of plants. Meanwhile, the invention also provides a method for producing an aluminum-resistant transgenic plant by using the gene, a kit for detecting the gene or protein and a method for carrying out assisted breeding by using the molecular marker linked with the gene. The invention provides a key gene resource for aluminum-resistant molecular breeding of crops, and has important significance for cultivating new varieties of crops adapting to acid soil.

The invention discloses recombinant IV-type collagen as well as a preparation method and application thereof, and belongs to the technical field of bioengineering. The amino acid sequence of the recombinant IV type collagen comprises a plurality of repetitive units, and the repetitive units are shown as SEQ ID NO. 1. The recombinant IV type collagen fragment is derived from human IV type collagen, heterologous amino acid residues are not introduced, and rejection and sensitization risks are avoided; meanwhile, the hydrogel also has good biological activity, including cell proliferation promoting activity and cell adhesion promoting activity, and is good in biocompatibility. In addition, the preparation method of the recombinant IV-type collagen is simple and easy to operate, and the recombinant IV-type collagen with high purity and stability can be obtained through the fermentation and purification method. Therefore, the recombinant IV type collagen disclosed by the invention has a relatively good application prospect.

The invention belongs to the technical field of bioengineering, and relates to a transaminase mutant, a recombinant genetically engineered bacterium and application of the transaminase mutant in catalytic synthesis of (R)-1-Boc-3-aminopiperidine.The transaminase mutant is obtained by conducting single-point or combined mutation on the 131 site and / or the 197 site of an amino acid sequence shown in SEQ ID NO.2; the mutation sites comprise that the 131 phenylalanine is mutated into aspartic acid, threonine or tyrosine, and / or the 197 lysine is mutated into arginine or leucine. Experimental results show that compared with wild type transaminase, the catalytic activity, the thermal stability and the organic solvent tolerance of the obtained mutants, especially single-point mutants MyTA1-F131Y and MyTA1-K197R and a combined mutant MyTA1-F131Y-K197R, are all remarkably improved, and compared with the wild type transaminase, the catalytic activity, the thermal stability and the organic solvent tolerance of the obtained mutants are all remarkably improved. The mutant MyTA1-F131Y-K197R can be used for efficiently catalyzing asymmetric amination of N-Boc-3-piperidone to synthesize (R)-1-Boc-3-aminopiperidine, the conversion rate of the (R)-1-Boc-3-aminopiperidine after the (R)-1-Boc-3-aminopiperidine reacts for 24 hours under the condition that the substrate concentration is 100 g / L can reach 90% or above, and the mutant MyTA1-F131Y-K197R has a good industrial application prospect.

The invention belongs to the technical field of biosynthesis, and particularly relates to an engineering bacterium for preparing stevioside through de novo fermentation as well as a preparation method and application of the engineering bacterium. According to the invention, mevalonate kinase in the MVA pathway is mutated and optimized. The method comprises the following steps: carrying out point mutation on mevalonate kinase MvK of a wild type source, constructing a stevioside synthesis route in an escherichia coli host, and comparing the influence of MvK mutation on the yield of stevioside, so as to determine a Q160L mutant, and applying the Q160L mutant to the stevioside production route to realize efficient production of stevioside.

The invention discloses a method for preparing butanediamine and spermidine from whole cells, and belongs to the field of bioengineering. According to a rational copolymerization modification strategy, a meridian interface and a latitudinal channel interface of arginine decarboxylase are subjected to rational modification respectively or simultaneously, and a series of mutants are obtained. The optimal mutant AdiA H729D / E467K / H736E, which is subjected to double-interface synergistic modification, disclosed by the invention, keeps high activity in the whole neutral-alkaline range of pH (Potential of Hydrogen) of 7.0 to 9.0. The optimal pH of the arginine decarboxylase mutant is increased, the enzyme activity stability in a neutral pH range is improved, the arginine decarboxylase mutant is more suitable for the condition requirements of industrial microbial fermentation, and a foundation is laid for efficient synthesis of butanediamine. The spermidine yield is up to 153.3 mg / L by using the mutant to catalyze the whole cell.

The invention discloses a method for preparing butanediamine and spermidine by an enzyme method, and belongs to the field of bioengineering. The novel arginine decarboxylase mutant is successfully provided, the mutant shows excellent catalytic performance under the alkaline condition, and the specific enzyme activity of the mutant reaches 24.9 U / g and is improved by 8.3 times compared with that of wild type enzyme. The invention develops a two-step synthesis process based on the constructed arginine decarboxylase mutant, and the two-step synthesis process is used for efficiently preparing spermidine. Experimental results show that the process successfully realizes the remarkable increase of the yield of spermidine, and the final yield reaches 200.4 mg / L.

The invention discloses application of a gene GlZn2Cys661 in regulation and control of synthesis of ganoderic acid. The nucleotide sequence of the gene GlZn2Cys661 is as shown in SEQ ID NO. 1. The gene expression is regulated and controlled by constructing an overexpression or silent vector, so that the synthesis of the ganoderic acid is promoted or inhibited, and an effective method is provided for regulating and controlling the yield of the ganoderic acid. The invention relates to the technical field of bioengineering, and discloses an application of a gene GlZn2Cys661 in regulation and control of ganoderic acid synthesis. The invention provides a key gene target and a core engineering strain for efficiently producing ganoderic acid through a metabolic engineering means, and has important application value.

The invention belongs to the technical field of bioengineering, and particularly relates to a biomass fermentation safety reinforcement learning control method and system, and the method comprises the steps: training an initial strategy network through imitation learning by employing historical data, and providing safety weight initialization for a reinforcement learning agent; and constructing a virtual training environment fusing the mechanism model and the data-driven residual error correction network, applying domain randomization, training an agent in the environment to maximize long-term accumulated rewards, and obtaining a robust control strategy. A training strategy is deployed to a real fermentation system, a control instruction is corrected in real time through an independent safety layer according to a hard constraint rule to ensure operation safety, and meanwhile online fine tuning and updating are conducted on the strategy based on actual operation data. According to the method, safe and smooth transition from virtual training to practical application is realized, and the final ethanol concentration and the overall control performance in the fermentation process are remarkably improved.

The invention belongs to the technical field of bioengineering, and particularly relates to a nucleic acid aptamer specifically bound with MMLV, and the nucleotide sequence of the nucleic acid aptamer is as shown in SEQ ID NO.1 or SEQ ID NO.2; the nucleic acid aptamer capable of being specifically bound with MMLV is obtained through screening on the basis of the SELEX technology, has high affinity for MMLV protein, can be applied to MMLV protein enrichment reagents, MMLV protein separation reagents, MMLV protein detection reagents, test paper and biosensors, and facilitates identification and activity research of the MMLV protein.

The invention discloses a melanin-producing aureobasidium for co-producing pullulan and converting American ginseng and application, and relates to the technical field of bioengineering fermentation. The melanin producing aureobasidium is aureobasidium melanogenum F8449, and the preservation number of the aureobasidium melanogenum F8449 is CGMCC (China General Microbiological Culture Collection Center) No. 23228. The strain can be used for simultaneously producing pullulan and biologically converting American ginseng extract in liquid, so that efficient utilization of fermentation substances is realized, the fermentation cost is reduced, and the fermentation income is increased.

The invention relates to the field of agricultural bioengineering, in particular to application of an oilseed rape BnaA08G0167600WE gene in enhancing pod shattering resistance of oilseed rape. According to the invention, after a transgenic plant is constructed in oilseed rape by utilizing a modern biotechnological means and the oilseed rape gene BnaA08G0167600WE is knocked out, the silique of the mutant is not easy to crack compared with a wild type under the action of external force, and a separation layer and a wood layer of the cross section of the silique of the mutant have more cell connections compared with the wild type, so that the yield of the mutant is improved. And the generation of the mutants provides an important genetic material for further researching the function of the gene and the effect of the gene in the rape.

The invention belongs to the technical field of bioengineering and application thereof, and particularly relates to a laver compound zymolyte as well as a preparation method and application thereof. The compound enzyme is prepared through a compound enzymolysis process of protease and laver polysaccharide enzyme. Compared with a traditional single proteolysis process, a composite enzymolysis process of adding the high-enzyme-activity laver polysaccharide enzyme effectively breaks a'polysaccharide-protein 'composite structure of laver cell walls, so that the viscosity of zymolyte is reduced, high laver liquefaction rate and high product yield are realized, and the release efficiency of nutritional active ingredients is remarkably improved; the product is rich in low-molecular-weight polypeptide and oligomeric porphyra polysaccharide. The obtained zymolyte shows the comprehensive effects of improving joint activity, inhibiting osteophyte formation, adjusting cartilage metabolism balance and relieving inflammatory response in an osteoarthritis mouse model, and osteoarthritis is effectively relieved. According to the invention, efficient release and utilization of nutritional active ingredients of the laver are realized, and powerful technical support is provided for developing high-valued products, such as health-care products and functional foods, which have the function of maintaining joint health.

The invention belongs to the technical field of biological engineering, and provides curcumin / dihydrocurcumin reductase MaCurA, a coding gene and application. The reductase can efficiently catalyze 1, 7-bis (4-hydroxy-3-methoxyphenyl)-1, 6-heptadiene-3, 5-diketone to synthesize tetrahydrocurcumin, an efficient solution is provided for industrial preparation of tetrahydrocurcumin, and experiments prove that the recombinase can catalyze the synthesis of tetrahydrocurcumin in the catalysis of 1, 7-bis (4-hydroxy-3-methoxyphenyl)-1, 6-heptadiene-3, 5-diketone. In a reaction system for preparing tetrahydrocurcumin from 2, 5-diketone, actually measured enzyme activity is stabilized at 3.38 U / mg level, and compared with a traditional chemical synthesis method, the production efficiency is remarkably improved.

The invention relates to the technical field of bioengineering, in particular to a preparation method of a probiotic and exosome preparation applied to immunoregulation. The probiotics and the umbilical cord stem cell exosome are combined for use, the immune regulation effect is enhanced from multiple targets and multiple ways, the immune function of the body is improved, the exosome carries a specific molecular marker and acts on immune cells in a targeted mode, precise immune regulation is achieved, and the preparation is in an oral form and convenient to use; the exosome is extracted from umbilical cord stem cells, has no pathogenicity and immunogenicity, and is low in adverse reaction risk after being used for a long time, and the prepared combined preparation can be used for treating and preventing various immune-related diseases, and also can be used for daily conditioning of people with low immunity and improving the body resistance; meanwhile, the combined preparation adopts micro-capsule embedding, so that the probiotics can be effectively protected from gastric acid and bile erosion, the survival rate and colonization efficiency of the probiotics in intestinal tracts are improved, the immunoregulation effect is further enhanced, and the combined preparation is convenient to store and transport and has a good application prospect.

The invention discloses a scallop skirt ACE inhibitory peptide activity method and system based on deep learning, and relates to the technical field of bioengineering.The scallop skirt ACE inhibitory peptide activity method comprises the steps that a comprehensive data set is obtained based on simulated in-vivo test data and peptide level feature data of different peptides, key features are screened, and a training set and a test set are divided; initializing a random forest model, an SVM model and a logistic regression model, performing independent training by using the training set, and determining an optimal prediction model of the peptide antihypertensive activity based on a prediction result of the test set; extracting importance data of key features based on the optimal prediction model for the antihypertensive activity of the peptide, sorting the importance data, and visually analyzing the antihypertensive activity features, feature importance sorting and model performance of different peptides; and all key results in the analysis process are integrated and systematically stored. According to the method, normal form reference with prediction accuracy and mechanism depth can be provided for clinical transformation of marine bioactive peptides, and AI-driven novel peptide drug research and development are promoted to enter a new stage.

The invention relates to the technical field of bioengineering, in particular to a penicillin G acylase mutant, polynucleotide, an expression vector and application. The penicillin G acylase mutant is obtained by carrying out site-directed mutagenesis on a wild type penicillin G acylase gene of parent Escherichia coli, and carrying out site-directed mutagenesis on the wild type penicillin G acylase gene to obtain the penicillin G acylase mutant. Phenylalanine (F) at the 24th site of an alpha chain, proline (P) at the 383rd site of a beta chain, threonine (T) at the 384th site of the beta chain, glutamic acid (G) at the 385th site of the beta chain and serine (S) at the 386th site of the beta chain of the penicillin G acylase are introduced and mutated by a whole plasmid PCR (Polymerase Chain Reaction) technology to obtain mutants. When the penicillin G acylase mutant obtained by the invention is used for catalyzing methyl mandelate and 3-(1-methyl-1H-tetrazole-5-yl) thiomethyl-7-aminocephalosporanic acid to synthesize cefamandole, the synthesis activity is improved, and meanwhile, the side reaction rate is greatly reduced.

The invention relates to the technical field of bioengineering, in particular to an exosome separation and concentration method with marine animal source tissue as a raw material. Comprising the following steps: freeze-drying and crushing a marine animal source tissue sample; performing sample enzymolysis; and removing impurities and concentrating. According to the method disclosed by the invention, the exosome in the cells is fully dissociated through freeze-dried powder crushing and composite enzymolysis processes, and the filtering speed of the sample is accelerated by virtue of ultrasonic oscillation, so that the rapid and high-purity separation and concentration of the exosome in the marine animal source tissue are realized.

The invention belongs to the technical field of bioengineering, and discloses a genetic engineering strain for enhancing schizochytrium limacinum DHA oil production based on carbon flux guidance and fatty acid transport, a method and application thereof. The gene engineering strain is obtained by introducing an optimized coding gene ACLY and / or Fat1p coding gene related to guiding carbon flux and fatty acid transport into a schizochytrium limacinum body; wherein the gene sequence of the gene ACLY is SEQ ID No. 9, and the gene sequence of the gene Fat1p is SEQ ID No. 10. According to the invention, a schizochytrium limacinum engineering bacterium for enhancing DHA oil production based on carbon flux guidance and fatty acid transfer is constructed by means of metabolic engineering. The schizochytrium limacinum engineering bacterium is combined with fatty acid transfer based on carbon flux guiding, can be used for grease synthesis by guiding carbon flux, and can prevent grease from being degraded through fatty acid transfer.

The invention belongs to the technical field of bioengineering, and discloses a high-yield hyaluronic acid bacillus subtilis strain for expressing a hyaluronic acid synthase gene, a construction method and application, and the construction method comprises the following steps: inserting a codon-optimized sphaasA gene into a lytH site in a bacillus subtilis 168 strain in a traceless manner by utilizing a CRISPR / dCas9 system; the preparation method comprises the following steps: synthesizing hyaluronic acid oligosaccharides, simultaneously integrating genes for expressing a synthetic pathway of hyaluronic acid oligosaccharides, reducing the transcriptional level of zwf and pfkA genes, weakening the metabolic flux of a competitive pathway, and finally increasing the expression of sphaasA and tuaD genes according to a ratio of 2: 1 by using plasmids and the interaction of protein scaffolds RIAD and RIDD. When the strain is fermented for 48 hours in a 5L fermentation tank, the yield of the hyaluronic acid reaches 38.02 g / L, and the average molecular weight HA Mw of the low-molecular-weight hyaluronic acid is about (5-13) * 10 < 4 > Da.

The invention relates to a biochemical method for preparing scopolamine, and belongs to the technical field of bioengineering. The invention aims to realize efficient conversion from scopolamine to scopolamine and improve the yield and industrial applicability of a target product. The invention provides a biochemical method for preparing scopolamine. The biochemical method comprises the following steps: adding scopolamine 6 beta hydroxylase into a culture medium containing L-scopolamine; the amino acid sequence of the hyoscyamine 6 beta hydroxylase is shown as any one of SEQ ID NO.1-5. The invention further discloses a preparation method of the hyoscyamine 6 beta hydroxylase. And the biochemical engineering method for realizing efficient and high-selectivity synthesis of scopolamine is realized.

A novel production scale-up adsorption separation device relates to the technical field of bioengineering product adsorption separation, and the technical scheme is: a filter is provided, the filter comprises an outer column pipe, an inner column pipe is arranged in the outer column pipe, and a hole is arranged in the inner column pipe; the filter is one or more than one; when the filter is one, a connecting piece is arranged in the middle of the outer column pipe, and N baffles are arranged on the connecting piece; the beneficial effects of the technical scheme are that the addition of the baffles enables the inner column pipe to form a relatively independent processing space, and liquid is circulated and communicated outside, so that the stability and uniformity of the fluid in the column pipe during the processing process are ensured, the production scale is expanded, the advantages of the original single equipment are guaranteed, the production land is saved, a plurality of single equipment is connected in series, each step is coordinated, the production efficiency is greatly improved, the utilization efficiency of the equilibrium liquid, the eluent and the like is improved, the cost is saved, and long-time automatic operation is realized.

The invention relates to the technical field of bioengineering, in particular to a preparation process for extracting antihypertensive peptide from euphausia superba. The process comprises the following steps: adding a ferulic acid / arginine double-grafted pullulan compound in a preparation process, and breaking an emulsification interface by utilizing beta-cyclodextrin adsorption and phospholipase A2 cracking to obtain a soluble pretreatment product; meanwhile, short-time round-trip pH pulse is implemented in the incision enzyme hydrolysis process to activate potential enzyme cutting sites, finally, hydrophobic aggregation peptides are removed through weak acid rapid elution, and excision hydrolysis is performed to obtain a target product with the molecular weight of 0.5-3 kDa, so that dynamic induction and antioxidant steady-state dual control of hydrolysis reaction is realized; the generation proportion of short peptides and the ACE inhibition persistence are improved, and the flavor and the storage stability are remarkably improved. The method is controllable in technological process, high in repeatability and suitable for large-scale preparation of the euphausia superba protein functional peptide and development and application of antihypertensive food.

The invention belongs to the technical field of bioengineering and microbiomics, and particularly relates to an acquisition method and application of a specific metabolic substrate of lactobacillus crispatus and application of a specific carbohydrate active enzyme CAZzyme gene and a polysaccharide utilization site (PUL). Genomes of lactobacillus crispatus and lactobacillus inertus are obtained from a public database, gene annotation is carried out on the genomes of the lactobacillus crispatus and the lactobacillus inertus, and a random forest model and a KEGG metabolic pathway diagram are combined to find specific CAZzyme genes, PUL and other genes with metabolic functions of the lactobacillus crispatus; a substrate which only lactobacillus crispatus can metabolize is searched based on the genome difference of the two bacteria, and the ability of the obtained substrate for promoting the growth of lactobacillus crispatus and inhibiting the growth of inert lactobacillus is evaluated through experiments. Growth and reproduction of the lactobacillus crispatus are promoted through the substrates, so that the lactobacillus crispatus accounts for a higher proportion in vagina microecology, the vagina is protected, and reproduction of other unfavorable bacteria is reduced. The assisted reproduction success rate is improved.

A bioengineering method which comprises introducing an inert nucleic acid cassette into a biological entity without introducing or modifying characteristics or traits in the biological entity. The method comprises receiving or providing a sample comprising the biological entity having a nucleic acid sequence; selecting an integration site in the nucleic acid sequence for inserting the inert nucleic acid cassette; designing the inert cassette with optimized primer sequences, optimized probe sequences, optimized stop codons and disrupted start codons, and inserting the inert nucleic acid cassette into the biological entity at the integration site; and validating that no characteristics have been added or modified in the biological entity.

The utility model discloses a kind of composite enzyme fermentation devices for bioengineering, it is related to composite enzyme fermentation technical field, solve the technical problem that the current reaction kettle in stirring process, single stirring rod drives stirring blade to rotate, and then make that material cannot be fully mixed, reduce the fermentation rate of composite enzyme, including stirring box, the feeding pipe is fixedly installed on the stirring box upper wall, the steam inlet pipe is fixedly installed on the stirring box side wall, the steam exhaust pipe is fixedly installed on the other side wall of stirring box, the utility model is mixed evenly by longitudinal stirring of stirring assembly to material, improve the fermentation rate of composite enzyme, by steam inlet pipe and steam exhaust pipe import hot steam, and then heat inner bag, assist composite enzyme to ferment, and then heat fermentation for material, by shock-absorbing component shock-absorbing protection to shock-absorbing box, provide stable support when material fermentation.