100 results about "Trichoderma reesei" patented technology

The invention discloses a special organic fertilizer for soybeans and a preparation method thereof, and belongs to the technical field of organic fertilizers. The special organic fertilizer for soybeans is prepared from the following raw materials in parts by weight: 8-10 parts of fish protein modified charcoal, 40-50 parts of decomposed soybean meal, 20-25 parts of decomposed livestock and poultry manure, 15-20 parts of humic acid, 10-15 parts of plant ash, 3-5 parts of a seaweed extract and 0.1-0.3 part of ammonium molybdate. According to the organic fertilizer, the soybean meal fermented by the trichoderma reesei and the aspergillus awamori and the fish protein modified biochar are added, the two components are compounded with the cooked livestock and poultry manure, the humic acid, the plant ash, the seaweed extract and the ammonium molybdate to form the organic fertilizer, the nitrogen fixation capacity of the soybeans is effectively improved through the synergistic effect of various substances, and the growth and development of the soybeans are remarkably promoted; the soybean yield and quality are improved. In addition, the whole organic fertilizer preparation process is free of strong acid and strong alkali treatment, the environmental pollution risk is reduced, and the standard of organic agriculture is met.

The invention discloses a method for screening high-yield recombinant human lactoferrin through nucleic acid aptamer-assisted mutagenesis of a trichoderma reesei strain and application, and belongs to the technical field of microorganisms. The nucleic acid aptamer Lac-6a-FAM-BHQ1 capable of changing the fluorescence intensity according to the content of lactoferrin is constructed on the basis of the aptamer Lac-6a, and the nucleic acid aptamer can be used for rapidly selecting a mutant strain for producing high-titer recombinant human lactoferrin. The ARTP mutagenesis technology is combined with a nucleic acid aptamer auxiliary screening system, the mutant strain with the improved recombinant human lactoferrin yield is rapidly obtained from a large number of mutant strains, and the yield reaches 274.57 mg / L and is improved by 24.8% compared with that of an original strain. After the genes 111681 and 62716 are over-expressed in T. reesei, compared with a control strain, the yields of the recombinant human lactoferrin of the recombinant strain are respectively increased by 6.9% and 22.1%.

The invention discloses a Trichoderma reesei strain and an application of the Trichoderma reesei strain in degradation of mycotoxin and inhibition of toxin-producing bacteria. The Trichoderma reesei strain is a Trichoderma reesei strain GG-T40, and the preservation number is CCTCC (China Center for Type Culture Collection) NO: M20231843. The strain can synchronously and efficiently degrade eight kinds of mycotoxins such as AFB1, ZEN, T-2, ST, OTA, AOH, AME and TeA, and after the strain and the mycotoxins are co-cultured for 7 days, the removal rates of the eight kinds of toxins are all larger than or equal to 60%. The strain has strong bacteriostasis and toxin production inhibition effects, the toxin production inhibition rates of the strain to aspergillus flavus and fusarium graminearum are 62.5% and 74.4% respectively, and the strain can be applied to degradation of mycotoxin in agricultural products and food and inhibition of mildew and toxin production of the agricultural products and food.

In the present application, by knocking out the BG gene in Trichoderma reesei PX3 (a strain of cellulase transcription activator that can be activated continuously), the BG activity can be reduced under the condition of not reducing or even increasing the filter paper enzyme activity of the enzyme system, and a low BG enzyme activity and high filter paper enzyme activity strain is constructed. This strategy can avoid the negative impact on cellulase production after BG knockout, and the obtained strain can be used for efficient production of cellobiose or as a chassis strain for producing cellulase.

The invention belongs to the field of fungus secondary metabolites and application thereof, and particularly relates to trichoderma reesei HK1-12 as well as a secondary metabolite and application thereof, the collection unit is China General Microbiological Culture Collection Center (CGMCC), and the collection number is CGMCC No. 41951. The trichoderma HK1-12 has excellent characteristics in the aspects of growth promotion, plant protection and the like, and is expected to replace the existing similar products to be popularized and applied.

The invention discloses an efficient composting and resource utilization method for garden waste. The method comprises the following steps: S1, classification pretreatment: classifying the garden waste into high lignin, medium lignin and low lignin; respectively crushing until the particle size is less than or equal to 5mm (class A) or less than or equal to 3mm; adjusting the initial carbon-nitrogen ratio (C / N) of the mixed material to (35-45): 1 and the initial water content to 55-65%; s2, preparation and inoculation of a complex microbial inoculant: preparing a complex functional microbial inoculant containing a lignin degrading microbial inoculant, a cellulose / hemicellulose degrading microbial inoculant, a function enhancing microbial inoculant and a protective carrier, and inoculating the complex functional microbial inoculant into the pretreated material with the inoculation amount being 0.5-1.0% of the total dry weight of the material. The garden waste is finely classified and crushed according to the lignin content, and the physical structure and the initial C / N of the material are optimized; the white rot fungi, the trichoderma reesei, the bacillus subtilis and other strains in the complex microbial inoculant have a synergistic effect, and particularly, the white rot fungi specifically degrade lignin, so that the degradation bottleneck of a lignocellulose composite structure is effectively broken.

The invention relates to a method for splitting, assembling and site-specific integration of large-fragment exogenous DNA (deoxyribonucleic acid) in Trichoderma reesei. Comprising the following steps: (1) constructing an arg:: Ptcu-Cas9-TtrpC-hph strain containing a Cas9 protein; (2) constructing a pUC19 <-> integration site: a pyr4 plasmid; (3) obtaining a plurality of target gene segments containing integration site homologous arms and having overlapping regions at the tail ends of adjacent DNA segments; (4) obtaining a gRNA expression cassette of a targeted genome integration site; (5) arg conversion: the arg is converted into a Ptcu-Cas9-TtrpC-hph strain; and (6) screening and verifying. According to the method, the CRISPR / Cas9 system is combined with the TAR in the trichoderma reesei for the first time, the exogenous large-fragment DNA is split into a plurality of small fragments, then the plurality of small-fragment exogenous DNA are assembled in the trichoderma reesei and are subjected to fixed-point integration, one-step editing of the large-fragment exogenous DNA is realized, and compared with a traditional genetics method, the gene editing efficiency in the trichoderma reesei is greatly improved.

This invention discloses a gene editing method and application for *Trichoderma reesei* based on a ribonucleoprotein complex. It aims to overcome the problems of low editing efficiency, easy generation of heterozygotes, and risk of exogenous DNA integration in *Trichoderma reesei* using plasmid-based CRISPR / Cas9 systems. This invention combines a dual-site cleavage strategy guided by double sgRNAs with direct RNP delivery. The method involves: designing two guide RNAs spaced 50-700 bp apart for the target gene, pre-assembling them with Cas9 protein in vitro to form an active RNP complex; delivering this complex along with exogenous donor DNA to *Trichoderma reesei* protoplasts. The exogenous donor DNA contains a left homologous arm, an editing sequence, and a right homologous arm, wherein the left and right homologous arms are homologous to the genomic sequences outside the dual cleavage sites on the target gene determined by the double sgRNAs, respectively; upon entering the cell, the RNP complex introduces dual-site cleavage at the target site, achieving precise editing under the guidance of the exogenous donor DNA.

The invention discloses a method for preparing a fermented feed from pomace and crop straw, and relates to the technical field of feed processing, and the method comprises the following steps: raw material treatment: performing rubbing treatment on corn straw; performing crushing treatment on the apple pomace; aerobic fermentation: uniformly mixing the filamentous material with the pomace powder, spraying a trichoderma reesei liquid into the mixture, and fermenting for 3-5 days under an aerobic condition; pressing and forming: after the aerobic fermentation is completed, pressing the mixed material into a cake belt with the thickness of 4-6mm through a double-roller tablet press; anaerobic fermentation is conducted, specifically, composite bacteria liquid is sprayed to the upper side and the lower side of the cake belt, the spraying amount of each square meter of the cake belt is 100-150 mL, and the composite bacteria liquid is composed of lactobacillus plantarum and saccharomyces cerevisiae according to the ratio of 3: 1; stacking the cake belt layers in a staggered manner; and feeding the cake belt into fermentation equipment, and fermenting for 2-3 days under an anaerobic condition. According to the method for preparing the fermented feed from the pomace and the crop straw, the feed is more balanced in nutrition, easy to digest and absorb and convenient for continuous fermentation production, and the fermentation product is convenient to store and transport.

The application discloses a recombinant Trichoderma reesei strain TriY120601 and a construction method and application thereof. Rut‑C30 The application discloses a recombinant Trichoderma reesei strain TriY120601 and a construction method and application thereof. Rut‑C30 The application discloses a recombinant Trichoderma reesei strain TriY120601 and a construction method and application thereof. Rut‑C30 The application discloses a recombinant Trichoderma reesei strain TriY120601 and a construction method and application thereof. Rut‑C30 The application discloses a recombinant Trichoderma reesei strain TriY120601 and a construction method and application thereof. Rut‑C30 The application discloses a recombinant Trichoderma reesei strain TriY120601 and a construction method and application thereof. Rut‑C30 The application discloses a recombinant Trichoderma reesei strain TriY120601 and a construction method and application thereof. Rut‑C30 The application discloses a recombinant Trichoderma reesei strain TriY120601 and a construction method and application thereof. Rut‑C30 The application discloses a recombinant Trichoderma reesei strain TriY120601 and a construction method and application thereof. Rut‑C30 The application discloses a recombinant Trichoderma reesei strain TriY120601 and a construction method and application thereof. Rut‑C30 The application discloses a recombinant Trichoderma reesei strain TriY120601 and a construction method and application thereof.

The present application provides a kind of shrimp shell biomass charcoal prepared by composite preparation and its preparation method and application, the present application is first prepared and obtained the composite preparation containing chitinase, bacillus licheniformis and bacillus subtilis, the chitinase is the chitinase derived from hartz wood enzyme is mutated, constructs mutant library, with trichoderma reesei as expression host, by screening, the excellent mutant Chi M1 obtained, its chitinase activity is significantly improved, is 2 times of original gene. By spraying the fermentation broth of the recombinant bacteria of the present application to obtain enzyme spray powder and the compounding of bacillus licheniformis powder and bacillus subtilis powder, the degradation efficiency of procambarus clarkia shrimp shell can be effectively improved. The composite preparation containing ChiM1 enzyme spray powder has good application prospect in the fields of aquaculture, feed additive and biomass resource utilization.

The application discloses a preparation method of small molecule peptides based on synergistic degradation of chicken eggshell membranes by molds and the small molecule peptides, and relates to the field of small molecule peptide preparation. The method comprises the following steps: mixing pretreated chicken eggshell membranes with mold fermentation liquor, and performing synergistic degradation; inactivating and centrifuging the degraded mixture, and collecting supernatant; purifying small molecule peptides in the supernatant through membrane separation technology; drying the purified small molecule peptide solution through a low-temperature spray drying method to obtain small molecule peptide powder; and the molds include Trichoderma reesei and Aspergillus niger. Through synergistic degradation of the two molds, the conversion of the chicken eggshell membranes is more thorough, the yield of the small molecule peptides is significantly improved, the total nitrogen recovery rate can reach 85.2%, and the product molecular weight distribution is concentrated.

The present application belongs to the technical field of nutritional functional food and health food, and particularly relates to a preparation method of marine fish skin collagen oligopeptide beneficial to promoting skin health, and a product and application thereof. The method comprises the following steps: after deep sea fish skin is pretreated by salt dissolution with a NaCl solution, trisodium citrate is added as an enzymolysis aid, and neutral protease (from Trichoderma reesei) and papain are sequentially used for two-step enzymolysis, and after enzyme inactivation, impurities are removed, sterilization is performed, and concentration, drying are performed to obtain the product. In vitro cell experiment verification shows that the product can significantly promote human skin fibroblast proliferation; in vivo animal experiment verification shows that the product can significantly increase skin moisture content, reduce transdermal water loss, increase hydroxyproline content, improve SOD activity and reduce MDA content. The product is suitable for the field of nutritional functional food and health food, and has a good application prospect in promoting skin health.

The invention discloses a method for preparing an amino acid fertilizer by multi-strain synergistic starter propagation and a liquid enzymolysis method, which comprises the following steps: selecting 25%-35% of bacillus subtilis, 10%-20% of trichoderma reesei, 15%-25% of aspergillus oryzae, 10%-20% of lactic acid bacteria, 5%-15% of saccharomyces cerevisiae and 5%-15% of aspergillus niger as composite strains; according to the method, six strains such as bacillus subtilis and trichoderma reesei form a composite microbial system, soybean meal and bran are used as raw materials, and the problems of low degradation efficiency, unstable enzyme activity and the like of a traditional process are solved through a process of'three-stage temperature-controlled starter propagation and four-stage liquid fermentation '; the method is characterized in that a'bacillus-mould-saccharomycetes-lactic acid bacteria 'four-flora synergistic system is constructed, a starter propagation-fermentation multi-parameter coupling regulation and control process is initiated, a'process-quality-effect' closed-loop mechanism is established, compared with a traditional process, the efficiency is improved by 35%-50%, the ton cost is reduced by 15%-20%, and the product is suitable for seed treatment and foliage spraying of various crops and has wide application prospects. The industrialization potential is remarkable, and the requirements of workers are met.

The invention belongs to the technical field of organic fertilizers, and particularly relates to a straw-based biochar composite slow-release fertilizer for saline-alkali soil improvement and a preparation method of the straw-based biochar composite slow-release fertilizer. The preparation method comprises the following steps: (1) uniformly mixing sheep manure and pigeon manure, mixing the mixed manure with crushed corn straw to obtain a total material, inoculating bacillus subtilis, and composting and fermenting to obtain a primary organic matter; (2) fermenting the primary organic matter by using aspergillus niger, trichoderma reesei and saccharomyces cerevisiae to obtain a submerged fermentation organic matter; (3) crushing the submerged fermentation organic matter to prepare particles; the composite coating powder is scattered into the submerged fermentation organic matter, coating is conducted, and the coated organic matter is obtained; and (4) uniformly mixing the coated fermented organic matter, biochar, desulfurized gypsum, humus soil, a water-retaining agent and ammonium dihydrogen phosphate to obtain the straw-based biochar composite slow-release fertilizer for saline-alkali soil improvement. The organic fertilizer can improve the pH value of saline-alkali soil, reduce the salt content and increase the organic matter content.

The present application relates to the technical field of microbial degradation of rhizoma polygoni multifidi residues, and discloses a composite microbial agent and a preparation method and application thereof.The composite microbial agent contains trichoderma reesei, aspergillus niger, trichoderma longibrachiatum, bacillus amyloliquefaciens, bacillus velezensis and bacillus subtilis; wherein the proportion of the spores or spores of the trichoderma reesei, aspergillus niger, trichoderma longibrachiatum, bacillus amyloliquefaciens, bacillus velezensis and bacillus subtilis in the composite microbial agent is 1-6:1-6:1-6:1-6:1-6:1-6:1-6.Based on this, the composite microbial agent has good degradation efficiency on rhizoma polygoni multifidi residues, can accelerate the composting process of rhizoma polygoni multifidi residues, shorten the composting period of rhizoma polygoni multifidi residues, and the composting product after composting is stable in quality and is a high-quality organic fertilizer, so that effective utilization of resources is realized.

The invention provides an efficient microbial fermentation inoculant for straws and a preparation method thereof, and relates to the technical field of biodegradation. The efficient microbial fermentation inoculant for the straw is formed by compounding four strains including trichoderma reesei, streptomycete, lactobacillus plantarum and bacillus amyloliquefaciens according to the mass ratio, the ratio of the viable count of the four strains is that the ratio of the viable count of the trichoderma reesei to the viable count of the streptomycete to the viable count of the lactobacillus plantarum to the viable count of the bacillus amyloliquefaciens is 3: 2: 2: 1.5, the total viable count of the inoculant is larger than or equal to 1.3 * 10 CFU / g, and the cellulase activity is larger than or equal to 85 U / g. The laccase activity is greater than or equal to 42U / g. The invention can effectively solve the industrial pain points of difficult degradation of straws, poor quality and weak stability, and in agricultural production, the microbial inoculum can increase the content of organic matters in soil, can reduce the feed cost in the feed field, and can be used for production of straw-based organic fertilizers in the environmental protection field, and can reduce the use amount of chemical fertilizers.

The invention discloses a laccase gene expression cassette, a laccase optical biosensor platform and a construction method and application of the laccase optical biosensor platform. The expression cassette comprises the following expression elements which are connected in sequence: (a) a Trichoderma reesei-derived Pcbh1 promoter; (b) an exogenous laccase coding gene; and (c) a Tcbh1 terminator derived from Trichoderma reesei. The invention has the following beneficial effects: the AALB platform realizes sensitive, economical and efficient screening of transcription factor variants by constructing a laccase gene expression cassette which is derived from oyster mushroom and is regulated by a Pcbh1 promoter; according to the invention, the high-activity exogenous laccase gene is integrated at the ACE1 site of the Trichoderma reesei genome, so that the regulation effect of the transcription factor is amplified, and the expression level of laccase can intuitively reflect the transcriptional activity of the transcription factor.

The present application relates to the technical field of microorganisms, in particular to a strain of Trichoderma reesei SDU1010, spore suspension and application, the Trichoderme reesei strain SDU1010 is preserved in the China General Microbiological Culture Collection Center on April 7, 2026, and the preservation number is CGMCC No. 3.29531.The Trichoderma reesei strain SDU1010 provided in the present application can effectively produce cellulase under acidic conditions with pH value of 2.5-5.5, effectively makes up the defect of low activity of key enzyme components under acidic conditions of existing strains, and improves the synergistic efficiency of cellulose degradation.

The invention discloses a preparation method and application of a composite bioactive additive for improving anaerobic fermentation efficiency, and belongs to the technical field of biomass energy. The preparation method comprises the following steps: constructing a mixed bacteria solid state fermentation system by using coriolus gallica NCULAC F1 (Coriolopsis gallica) CCTCC M2021731 with high laccase yield and Trichoderma reesei SHBCC D23843 with high cellulase yield, and preparing the composite bioactive additive by virtue of regulation and control of an enzyme production induction liquid and a two-stage solid state fermentation process, so as to obtain the coriolus gallica / Trichoderma reesei / Trichoderma reesei / Trichoderma reesei / Trichoderma reesei / Trichoderma reesei / Trichoderma reesei / Trichoderma reesei composite bioactive additive. The composite bioactive additive is used for pretreatment of straw stalks, the yield of biogas and methane can be remarkably increased, and the fermentation period is shortened.

The application discloses a kind of anti-saline-alkali microbial inoculants and preparation method thereof, belong to the field of microbial technology.The microbial inoculants is made of composite microbial inoculants 100-150 parts, fish bone hydrolysate 10-30 parts, nano silicon sol 1-3 parts.The composite microbial inoculants is made of Trichoderma reesei, Pseudomonas plecoglossicida, Acinetobacter baumannii.The anti-saline-alkali microbial inoculants has remarkable effect on saline-alkali soil improvement, greatly reduces soil acidity, salt content and bulk density, and the yield of saline-alkali soil wheat planting is obviously improved.The soil organic matter content and soil enzyme activity are significantly improved after use, the soil quality and rhizosphere microecological environment are improved, and the enzyme activity in wheat is improved, the stress resistance is improved, the yield reduction caused by saline-alkali stress is reduced, and the use of microbial inoculants can also reduce the amount of chemical fertilizer and pesticide, reduce production cost, and further improve the economic benefit of wheat planting.

The invention is suitable for traditional Chinese medicine residue resource utilization and biological refinery technology, and provides a preparation method of a high-protein and high-fat feed. The method comprises the following steps: adding a certain amount of cheap nitrogen source into plant source wet traditional Chinese medicine residues rich in starch to prepare a fermentation culture medium, inoculating trichoderma reesei and lipomyces starkeyi for mixed culture, and performing combined action on cellulase produced by the trichoderma reesei and amylase produced by the lipomyces starkeyi on the traditional Chinese medicine residues, so as to obtain a fermentation product. And rapidly degrading into fermentable sugar which is efficiently used for microbial growth and grease and protein production, then carrying out solid-liquid separation, and drying the solid to obtain the high-protein and high-grease feed. Wet traditional Chinese medicine residues rich in starch are used as raw materials, cellulase and amylase production, enzyme hydrolysis and fermentation are highly integrated, the number of equipment is reduced, the production and processing flow is simplified, and the production cost of the high-protein feed is remarkably reduced. The fermented product serving as the feed is rich in protein and microbial oil, the fatty acid composition of the microbial oil is similar to that of vegetable oil, the nutritional ingredients of the feed are further improved, and rapid growth of poultry can be efficiently promoted. The method can promote the resource utilization of the cheap traditional Chinese medicine residues, provides a low-cost new method for improving the feed quality, and has remarkable economic benefits.

The application discloses a method for improving cellulase activity by using a combination of an endogenous promoter and an associated 5' UTR of Trichoderma reesei and an engineering strain constructed by the method. TrAE The 5' UTR upstream of the TrAE gene TrAE Overexpressing a transcriptional regulator xyr1 The expression cassette of the gene is transformed into Trichoderma reesei Δ ku70 Rut‑C30 The application also discloses a high-yield cellulase Trichoderma reesei strain QEaeuX, and a method for producing high-activity cellulase by using the strain. ku70 Rut‑C30 The cellulase activity produced by the strain in a 96 h shake flask culture is 2.6 IU / mL, which is 101% higher than that of the original strain Δ The application has a good industrial development and application prospect.

The invention relates to a strain of Trichoderma reesei BLCY-007 and its application in the production of xylooligosaccharides.

The application discloses a new Sorbicillinoids compound and an extraction and separation method and application thereof, relates to the technical fields of natural compound extraction, separation and natural medicine chemistry. The application takes Trichoderma reesei fermentation liquor as raw material, extracts a new Sorbicillinoids compound from Trichoderma reesei metabolites, and names the compound as Trichosorbicil. The molecular formula of the Sorbicillinoids compound Trichosorbicil is C 25 H 28 O7, and a structural formula is as follows: The compound Trichosorbicil has relatively significant in-vitro anti-tumor activity, and provides a basis for further developing in-depth pharmacological and clinical research and developing a new anti-tumor drug with high curative effect and small side effect.

The invention discloses a preparation process of a multi-stage fermentation biological feed, and belongs to the technical field of feed biology. In order to solve the problems that the existing fermented feed is poor in strain synergism, insufficient in enzymolysis, low in probiotic colonization amount and the like, the method adopts a staged and time sequence inoculation strategy, and four stages of aerobic enzyme production degradation, aerobic fermentation and enzymolysis, facultative anaerobic fermentation and anaerobic probiotic colonization are sequentially carried out; and the inoculation sequence and proportion of trichoderma reesei, aspergillus niger, aspergillus oryzae, bacillus subtilis, candida utilis and lactobacillus plantarum are accurately controlled. By constructing an efficient flora metabolic chain, the quality of the feed is remarkably improved. The crude protein content of the prepared feed can reach 37.8% or above, and the viable count of lactobacillus plantarum reaches 3.1 * 10 < 10 > CFU / g. The prepared feed product has the advantages of high nutritional value, strong probiotic function and long shelf life, and the process is controllable and suitable for industrial production.

The invention relates to a modified cellulase derived polypeptide and application thereof, and belongs to the technical field of gene engineering and enzyme engineering. The invention provides a derivative polypeptide of cellulose excision enzyme Cel7A, the cellulose excision enzyme Cel7A is derived from Trichoderma reesei, NCBI Acession No.XM006969162.1, and the amino acid sequence of the cellulose excision enzyme Cel7A is as shown in SEQ ID NO: 1. The invention further provides a preparation method of the derivative polypeptide of the cellulose excision enzyme Cel7A. Wherein the amino acid sequence of the cellulose binding domain CBM is as shown in SEQ ID NO: 2. The derivative polypeptide is a polypeptide of which the amino acid sequence is 70% or more identical to that of a polypeptide shown in SEQ ID NO: 2, can reduce combination with lignin and improve the degradation efficiency of cellulose, so that the saccharification efficiency of cellulase is improved, the post-treatment process cost is greatly saved, and the derivative polypeptide is suitable for industrial production. The method has important industrial application value.

The application discloses a kind of for decomposing forest litter composite microbial inoculant, preparation method and its application method, it is related to litter decomposition technical field.The application includes a kind of for decomposing forest litter composite microbial inoculant preparation method, comprising the following steps: S1.to Phanerochaete chrysosporium, Trichoderma reesei and ropegillus catenulatum are respectively carried out directional metabolism preactivation;S2.the three preactivation wet bacteria bodies obtained in step S1 are mixed according to the fresh weight ratio of Phanerochaete chrysosporium:Trichoderma reesei:ropegillus catenulatum 3:2:1, obtain mixed bacteria body;S3.mixed bacteria body is mixed with the straw carrier that is preprocessed by steam explosion according to the mass ratio of bacteria body fresh weight:carrier dry weight 1:2, under the condition of temperature 28 DEG C, relative humidity 92~95%, stationary culture 72 hours, carry out phased micro-perturbation aeration, construct physical symbiosis;S4.the material obtained in step S3 is dried to 15% moisture content at 40 DEG C, obtain composite microbial inoculant;Efficient and stable decomposition of pine forest litter is realized.

The invention discloses a method for preparing complete feed based on multi-strain fermentation, and belongs to the technical field of feed processing. Comprising the following steps: pretreating a raw material through cellulose compound enzyme, sequentially carrying out aerobic synergistic fermentation of aspergillus niger and trichoderma reesei and anaerobic compound fermentation of lactobacillus plantarum-saccharomyces cerevisiae-bacillus subtilis to realize deep degradation and nutrition conversion of crude fibers in the raw material, and finally preparing a fermentation product into the duck complete feed. The problems of incomplete degradation, high infectious microbe pollution risk and mutual inhibition among strains of a traditional fermented feed are solved, the nutritional value and the utilization efficiency of agricultural by-products are improved, the prepared feed is rich in probiotics, organic acid and small molecule nutrient substances, corn-soybean meal type feed can be replaced, grain conflict is reduced, and the production cost is reduced. Meanwhile, agricultural by-product waste pollution is reduced, economic and environmental benefits are achieved, and the method is suitable for large-scale industrial popularization.

The invention provides a Trichoderma reesei engineering bacterium for high yield of beta-glucosidase, which is characterized in that a Trichoderma reesei SN1 strain is used as a starting strain, and an expression cassette for knocking out a cre1 gene, a constitutive expression cassette of an aspergillus niger source bglA gene expressed by a promoter cbh1 and a constitutive expression cassette of a 130442 gene expressed by the promoter cbh1 are constructed in a genome of the Trichoderma reesei SN1 strain. Through the knockout of the cre1 gene, the overexpression of the aspergillus niger source bglA gene and the overexpression of the 130442 gene, the yield of the beta-glucosidase in the trichoderma reesei is improved synergistically. Glucose is taken as a raw material, a transglycosylation reaction can be catalyzed by directly utilizing a crude enzyme solution of trichoderma reesei engineering bacteria with high yield of beta-glucosidase to prepare beta-disaccharide, and the beta-disaccharide can be taken as an inducer for cellulase production; a crude enzyme liquid with higher cellulase activity and beta-glucosidase activity is obtained by supplementing an inducer and carrying out fed-batch fermentation.