34 results about "Thermomyces lanuginosus" patented technology

The invention discloses a high-temperature cow dung decomposition agent which is prepared from cow dung and a compound microbial agent through fermentation and drying, wherein cow dung particles in the high-temperature cow dung decomposition agent are loaded with compound bacteria of bacillus subtilis, bacillus licheniformis, in-situ composting geobacillus toebii, thermomyces lanuginosus and thermobifida fusca. In addition, the living bacteria count is not lower than 48+/- 100 millions/g, the high-temperature cow dung decomposition agent is 50-650 [mu]m in average particle size, lower than 15% in moisture content and 6.0-8.0 in pH. The compound microbial agent in the decomposition agent generates plenty of cellulase, hemicellulase and protease, so that cow dung is rapidly decomposed into a simple inorganic nitrogen source and a carbon source, further, plant growth is promoted and soil fertility is improved. Fermentation is carried out under the high temperature environment, so that mesophilic plant pathogen growth is inhibited effectively, and cow dung products fermented through the decomposition agent can be directly used as organic fertilizer to be applied to soil.

The invention discloses an organic fertilizer fermentation inoculant and application. The inoculant is composed of a primary fermentation inoculant body and a secondary fermentation inoculant body. The primary fermentation inoculant body comprises bacillus subtilis, bacillus licheniformis, trichoderma viride, aspergillus oryzae, pseudomonas, trichoderma koningii, geotrichum candidum, long spiny shells, ascomycetes, thermomyces lanuginosus and thermobifida fusca. The secondary fermentation inoculant body comprises halophilic bacteria, alkalophilic bacteria, azotobacter chroococcum, bacillus megatherium, streptomyces griseus, bacillus thuringiensis, brevibacillus laterosporus and bacillus mucilaginosus. Time for fermenting organic fertilizer with cow dung as the raw material can be effectively shortened, and diseases and insect pests in the fertilizer can be eliminated. Meanwhile, alkalophilic bacteria and bacillus thuringiensis are added so that the fermentation inoculant can adapt to saline-alkali soil and can prevent diseases and insect pests of plants, and the prepared organic fertilizer can improve the soil structure, reduces the salt content and the pH value in soil, improves soil fertility and is particularly beneficial to improvement of saline-alkali soil.

The invention discloses a maize straw high-temperature decomposition agent. The maize straw high-temperature decomposition agent is prepared by fermenting and then drying maize straw powder and complex microbial inoculants. The maize straw powder particles in the maize straw high-temperature decomposition agent are loaded with a bacterium mixture containing thermomyces lanuginosus, thermobifida fusca, thermopolyspora flexuosa, streptomyces thermoviolaceus, in-situ composting bacillus licheniformis and bacillus subtilis. Living bacteria count is not lower than 48+/-1 hundred million/g. The average grain diameter of the maize straw high-temperature decomposition agent is 150-650 micrometers, water content is lower than 15%, and pH is 6.0-8.0. By the adoption of the decomposition agent, the lignocelluloses substance and the protein substance in maize straw can be degraded quickly to achieve straw decomposing, and rich simple inorganic carbon sources and nitrogen sources are generated. Decomposed maize straw is made into organic fertilizer to effectively promote the growth of plants and improve the fertility of soil. Soil improvement and environment management are facilitated, and sustainable development of agriculture is achieved.

The invention discloses a thermomyces lanuginosus lipase gene, engineering bacteria and an application of the engineering bacteria. The base sequence of the thermomyces lanuginosus lipase gene is shown in SEQ ID NO.2. On the basis that existing thermomyces lanuginosus lipase amino acid sequences and protein structures are not changed, factors adverse to protein expression, for example, rare codons, rare codon pairs, strong stem-loop structures, potential early termination signals and the like are avoided, and the thermomyces lanuginosus lipase gene sequence is optimized, so that the sequence can efficiently express thermomyces lanuginosus lipase in a pichia pastoris recombination expression system, and the expression vitality is up to 370.65 U/ml and is improved by 58.38% while compared with the recombination expression vitality of the original sequence of the thermomyces lanuginosus lipase gene being 234.03 U/ml.

The invention discloses a method for immobilizing Thermomyces lanuginosus lipase. The method comprises the following steps: adding 100 L of an enzyme liquid of Thermomyces lanuginosus lipase Lipozyme TL into an organic solvent at 4 DEG C, carrying out standing precipitation for 1 h, centrifuging the obtained solution, taking the obtained supernatant, adding a cross-linking agent and macro-porous adsorption resin, carrying out oscillation cross-linking immobilization, and carrying out vacuum drying after the above reaction is completed in order to obtain immobilized Thermomyces lanuginosus lipase. The immobilized lipase has the advantages of simple preparation conditions, and convenience in operation. The enzyme activity and the batch stability of the immobilized Thermomyces lanuginosus lipase are much higher than those of the commercialized enzyme Lipozyme TL IM, so the method is suitable for industrial application.

The invention provides a method for increasing the oligomerization wood sugar content in black tea aiming at the disadvantages that the oligomerization wood sugar content in the black tea at present is low and the effect is inadequately obvious. The method is characterized in that heat-resistant xylanase is produced in the black tea fermentation process through inoculating a xylanase producing strain, namely Thermomyces lanuginosus, and thus oligomerization wood sugar produced by xylan in the black tea is degraded; detection results prove that compared with the traditional black tea, the oligomerization wood sugar content in the black tea prepared by using the method is increased by 20-40 times; the animal experiment results prove that bifidobactirium proliferation in the intestinal tracts of white rats fed by the black tea is obvious and is improved by about 8% compared with that in the intestinal tracts of the white rats fed by the traditional feeding black tea, and the growth of enteric bacilli, enterococcus and the like can be inhibited to different degrees.

The invention discloses a preparation method of a thermomyces lanuginosus lipase. The method comprises the following steps of: 1) synthesizing a thermomyces lanuginosus lipase gene; 2) constructing a thermomyces lanuginosus lipase expression plasmid; 3) converting a trichodermareesei and selecting high-yield strains of the thermomyces lanuginosus lipase; and 4) separating and purifying the thermomyces lanuginosus lipase. The invention also discloses the application of the thermomyces lanuginosus lipase to the preparation of flour foods.

The invention provides a garden waste degradation bacteria agent. The garden waste degradation bacteria agent comprises a bacteria groupand a basic culture solution, wherein the bacteria group comprises Promicromonospora, Burkholderia, Thermomyces lanuginosus, Shigella and Streptomyces; the basic culture solution comprises wheat bran, starch, xylose, beef extract, ammonium nitrate and sterile water; the weight ratio of the bacteria group to the basic culture solution is 1:1.5-1:2. The garden waste degradation bacteria agent has the advantages that after fermentation of garden wastes is finished by using the degradation bacteria agent, the degradation rates of hemicellulose, cellulose and lignin are obviously higher than those obtained after one kind of bacteria is individually used; the Shigella and the Streptomyces are degraded by adding sugar, other polysaccharides in the garden wastes can be efficiently degraded, and the overall fermentation process is improved. The compound degradation bacteria agent is high in lignin degradation efficiency; land resources are saved, soil and the ecological environment are improved, environmental pollution is mitigated, the substrate obtained after degradation can be turned into treasure, and resource recycling is achieved.

The invention relates to a method for enriching long-chain polyunsaturated fatty acids through immobilized thermomyces lanuginosus. The method includes following steps: (1) carrying out an esterification reaction to a free fatty acid and an alcohol in the presence of the immobilized thermomyces lanuginosus in vacuum or a protective atmosphere; and (2) separating free long-chain polyunsaturated fatty acids out from a product after the reaction. By means of selectively catalytic esterification to fatty acids with different chain lengths during a catalytic esterification process of the fatty acid and the alcohol through immobilized lipases, a new method for enriching the long-chain polyunsaturated fatty acids from deep sea fish oil and microbial oil is achieved.

The invention provides a thermomyces lanuginosus lipase mutant, coding genes, and applications of the thermomyces lanuginosus lipase mutant. The thermomyces lanuginosus lipase mutant is obtained via point mutation of a peptide fragment with a sequence represented by SEQ ID No.1; and the point mutation is one selected from the following steps or a combination of the following steps: (1) S at the 63th site is changed into L or M; and (2) D at the 232th site is changed into A. Compared with the parent, vitality of the thermomyces lanuginosus lipase mutant is increased greatly; the thermomyces lanuginosus lipase mutant can be used for industrialized production of (3S)-2-carboxyethyl-3-cyano-5-methylhexanol, and further synthesis of pregabalin.

The invention discloses a method for preparing oligosaccharides with side chains by utilizing a thermophilic filamentous fungus fermentation one-step method. The method is characterized in that the oligosaccharides with the side chains are prepared by utilizing a method for solid fermentation or liquid fermentation of thermomyces lanuginosus. An experiment verifies that through the thermophilic properties of the thermomyces lanuginosus, fermentation conditions are easy to control in the production process of the thermomyces lanuginosus; meanwhile, the thermomyces lanuginosus can utilize cheapcorn cobs, bran and other agricultural and sideline products to ferment and produce the oligosaccharides with the arabinose side chains; the invention disclosed by the invention is a sustainable production method, can reduce the raw material cost and the separation and purification cost and is an environment-friendly and efficient production process.

The invention discloses an esterase, a gene, a vector and an engineering bacterium derived from thermomyces lanuginosus as well as application in preparing (3S)-2-carboxyethyl-3-cyan-5-methyl hexanoic acid. An amino acid sequence of the esterase is represented in SEQ ID NO: 2. The esterase gene disclosed by the invention can be linked to an expression vector to construct an intracellular expression recombinant plasmid containing the gene and the intracellular expression recombinant plasmid is transformed to an escherichia coli strain, thus obtaining recombinant escherichia coli. The recombinant escherichia coli contains the esterase; the recombinant escherichia coli or purified esterase is applied to biological catalysis as a catalyst to produce a pregabalin key chiral intermediate.

The invention provides a lipase-calcium phosphate compound enzyme crystal, a preparation method thereof and a method for catalyzing and synthesizing clindamycin palmitate by using the lipase-calcium phosphate compound enzyme crystal, and belongs to the technical field of antibiotic drug synthesis. According to the method, clindamycin free alkali shown in the formula I and ethenyl hexadecanoate shown in the formula II serve as a substrate and react with a reaction solvent for 6-30 h at the temperature of 30-80 DEG C under the catalytic function of the lipase-calcium phosphate compound enzyme crystal, and clindamycin palmitate shown in the formula III is prepared. The lipase-calcium phosphate compound enzyme crystal prepared through the method is a calcium phosphate crystal of thermomyces lanuginosus lipase. The lipase-calcium phosphate compound enzyme crystal has the advantages that the cost of the method is low, immobilized enzyme cannot be dissolved in an organic solvent and can be recycled, the yield can reach 75.8% at most, the method is safe, reliable, low in environment pollution and suitable for industrial production.

The invention discloses a lipase composition and application thereof, and belongs to the technical field of enzyme engineering. The lipase composition is composed of 25-50% of lipase from pseudomonasaeruginosa and 50-75% of lipase from thermomyces lanuginosus according to the protein content. The lipase composition has the synergic effect for a hydrolysis reaction and an esterification reaction for catalyzing grease. Triglyceride can be rapidly hydrolyzed, the esterification initial reaction speed is increased, the reaction process is more gentle, fluctuations are little, when the reaction isended, more esterification products can be generated, the enzymic preparation dosage is lowered, the reaction time is effectively shortened, the production cost is effectively saved, and the excellent application and popularization value is achieved in the fields of grease chemical industry, feed additives, detergents and the like.

The invention discloses a thermomyces lanuginosus lipase mutant and a coding gene. The amino acid sequence of the mutant is shown as SEQ ID NO: 2. According to the mutant disclosed by the invention, polar amino acid at a specific position is mutated into non-polar amino acid, site-directed mutagenesis molecular modification of rational design is carried out on existing enzyme molecules, the mutated enzyme molecular structure is not destructively changed, a mutation result is relatively safe, and enough enzyme activity can be ensured or higher enzyme activity can be obtained.