136 results about "Feruloyl esterase" patented technology

Ferulate esterase producing bacterial strains or functional mutants thereof and methods of using ferulate esterase producing bacterial strains as forage additives are disclosed.

The invention belongs to the technical field of bioengineering, and provides a feruloyl esterase high-producing strain and a method for preparing feruloyl esterase through liquid fermentation by using the same. The feruloyl esterase production strain provided by the invention is a filamentous fungus; 16S rDNA identification shows that the feruloyl esterase production strain is a Trichodermaviride; the strain code is JBSH-001; and the preservation number of China General Microbiological Culture Collection Center is CGMCC NO.5612. Feruloyl esterase can be produced through liquid fermentation by using the strain. The enzyme activity of the feruloyl esterase obtained through fermentation according to the method can reach 200 mU/ml or above; and the enzyme activity of the purified feruloyl esterase can reach 50000 mU/g or above.

The invention discloses a feruloyl-esterase-producing Bacillus licheniformis strain and application thereof. The collection number of the Bacillus licheniformis DBM12 strain is CGMCC No.8672. The Bacillus licheniformis strain has the advantages of lower requirements for the carbon source and nitrogen source, short fermentation time, high safety and no toxicity, and can be used in food/feed industry. The Bacillus licheniformis strain has favorable hereditary stability, and is simple to operate. The feruloyl esterase synthesis capacity is basically unchanged after serial passage by more than 10 generations.

The invention provides a feeding complex ferulic acid esterase additive, which comprises four enzymes of ferulic acid esterase, cellulase, xylanase and glucanase; among which, the enzyme activity of endo cellulose is more than 16U/g, the enzyme activity of xylanase is more than 1024U/g, the enzyme activity of beta-glucanase is more than 2.0U/g, and the enzyme activity of ferulic acid esterase is more than 2.0U/g. The feeding complex ferulic acid esterase additive of the invention has the advantages of remarkably improving the release amount of reducing sugar, the cell wall degrading rate and the vitro rumen fermentation volatile fatty acids yield in the hydrolysis process of forage plants, crop straws and bran feed, enhancing the feed biological transfer rate in an all-round way. The invention has a high practical application value in the feed industry and the ruminant breeding industry.

The present invention relates to an oral formulation to lower serum or hepatic lipid and triglyceride concentrations, hepatic inflammation and/or insulin resistance in a patient comprising live feruloyl esterase producing microorganisms alone or in association with a pharmaceutically acceptable carrier resistant to gastric conditions, and wherein the microorganisms are wild type, genetically modified, or combination thereof. The present invention is also directed to an oral formulation to lower serum or hepatic lipid and triglyceride concentrations, hepatic inflammation and/or insulin resistance in a patient, which comprises polymeric microcapsules containing live feruloyl esterase producing microorganisms in suspension in a pharmaceutically acceptable carrier, wherein said microcapsules are semipermeable and resistant to gastro-intestinal conditions, and wherein said microorganism are wild type, genetically modified, or combination thereof as well as methods of preventing or improving liver diseases and disorders and uses thereof.

The invention belongs to the technical field of gene engineering and enzyme engineering, specifically to a feruloyl esterase A mutant and the purpose thereof. According to the invention, parent genes of feruloyl esterase A undergo a random mutation by error-prone PCR to construct a random mutation library; 12 heat-resistance enhanced feruloyl esterase A mutant sites are selected by a high-throughput screening system; and a complete sequence synthetic method is adopted to integrate the 12 mutation sites onto a coding sequence so as to obtain a combination mutant. The combination mutant has much more enhanced heat stability than the parents and is of potential application value in industrial fields such as bioenergy, forage, papermaking, health food, medicine and the like.

The invention relates to a method for preparing xylo-oligosaccharide syrup and powdered sugar by using a complex enzyme. The method comprises the following steps of: (1) crushing a cellulosic raw material and preparing the cellulosic raw material into a suspension; (2) adjusting the pH value of the suspension to be between 5.5 and 6.5, adding a high temperature-resistant alpha-amylase into the suspension, after raising the temperature to 85 to 95 DEG C, hydrolyzing the suspension, cooling the suspension, adjusting the pH value to be between 7.5 and 8.5, adding an alkali protease, stirring the suspension at a constant speed, after raising the temperature to 45 to 60 DEG C, hydrolyzing the suspension, and inactivating the enzyme; (3) centrifuging the suspension in which the enzyme is inactivated to precipitate, and washing and drying the deposit; (4) preparing the deposit into a dietary fiber suspension, adjusting the pH value to 5.6, and adding the liquid complex enzyme consisting of incision beta-1,4-xylanase, feruloyl esterase and fungal laccase into the suspension to obtain a xylo-oligosaccharide enzymatic hydrolysate; and (5) performing precise filtration, nano filtration, super filtration, desalination, decoloration and concentration on the xylo-oligosaccharide enzymatic hydrolysate to obtain the xylo-oligosaccharide syrup. The method is simple in process and wide in raw material source; the raw materials can be mixed; the production time is shortened; the production efficiency and the yield are improved.

The invention provides feruloyl esterase and a preparing method and application thereof. A feruloyl esterase gene coming from a soil macro gene library have the nucleotide sequence and amino acid sequence shown in SEQ ID NO.1 and SEQ ID NO.2. The gene contains a tetrapeptide SXXK sequence motif which is rarely seen, and after the esterase gene is inserted into plasmid pET28a(+), the gene is transformed into escherichia coli BL21(DE3) to achieve heterogeneous expression. The molecular weight of purified recombinase (DLFae4) is 38.3 kDa. Besides, it is put forward for the first time that novel feruloyl esterase can hydrolyze penbritin, penicillin, cefazolin and other lactam antibiotics. As is shown by site-directed mutagenesis experiments, a catalysis triplet of DLFae4 is composed of serine(S11), histidine (H74) and aspartic acid (D302), and the mutation of any of serine (S11), histidine (H74) and aspartic acid (D302) can cause loss of the catalysis capability of DLFae4. DLFae4 has a high hydrolytic activity on methyl ferulate and has good heat stability. In the presence of cellulase, DLFae4 can obviously increase the amount of ferulic acid released from destarched wheat bran. Due to peculiar activities and enzymatic characteristics of novel feruloyl esterase, novel feruloyl esterase can be applied to feed, paper making, food, pharmacy and other fields.

The invention discloses Cladosporiumc cladosporioides for high yield of feruloyl esterase and an application of Cladosporiumc cladosporioides in edible vinegar brewing, and belongs to the field of fermentation engineering and biotechnology. The Cladosporiumc cladosporioides was preserved by the China Center for Type Culture Collection, the preservation number is CCTCC NO: M2015549, and the preservation date is September, 16th, 2015. According to the invention, wheat bran is used as a fermentation culture medium for fermentation production of high-activity feruloyl esterase, and it is easy to realize industrial production of feruloyl esterase. By applying feruloyl esterase to production of edible vinegar, functional health-care edible vinegar rich in ferulic acid is obtained. Thus, new technical support is provided for development of health-care vinegar.

The invention discloses a preparation method of recombinant feruloyl esterase. The feruloyl esterase O42807 is adopted, after codon optimization, the gene fae of the feruloyl esterase O42807 is manually synthesized, A pAO815-fae expression vector is established, a pPIC9K-fae expression vector is established and the pPIC9K-fae expression vector is transformed to pichia pastoris GS115 for expression, and then the recombinant feruloyl esterase is obtained. SDS-PAGE analysis shows that the fermentation liquid supernatant of the recombinant feruloyl esterase is a single stripe, the apparent molecular weight is 42kDa, the enzyme activity is 4.7U/mL, the enzymatic specific activity is 31.4U/mg, the optimal reaction temperature is 50 DEG C and the optimal pH is 5.0-5.5. According to the preparation method of the recombinant feruloyl esterase, an efficiently expressed recombinant feruloyl esterase transformant is obtained, and the expression efficiency and the expression quantity of the recombinant feruloyl esterase transformant are greatly improved in contrast with feruloyl esterase O42807.

The invention provides a feruloyl esterase gene. The feruloyl esterase gene is originated from microbes in the rumen of a Chinese yak and has a nucleotide sequence as shown in SEQ ID No. 1, wherein the nucleotide sequence codes an amino acid sequence as shown in SEQ ID No. 2. The invention also provides a recombinant Kluyveromyces marxianus expression vector containing the gene, a genetically engineered strain, a preparation method for feruloyl esterase and application of prepared feruloyl esterase. Feruloyl esterase is subjected recombinant expression in a Kluyveromyces marxianus expression system; and after high-density fermentation for 48 h, intracellular and extracellular feruloyl esterase vitalities are 686.35 U/mL and 346.34 U/mL, respectively. Feruloyl esterase having undergone recombinant expression by Kluyveromyces marxianus can release ferulic acid in corn bran. Feruloyl esterase prepared in the invention can be applied to food processing, feed addition, biomedicine, biotransformation of cellulosic substances, bioenergy and other fields.

The invention provides preparation of an enzyme preparation with acidic proteinase as a main component as well as a strain and application of the enzyme preparation. The multi-enzyme series acidic proteinase preparation which utilizes the acidic proteinase as the main component, utilizes multiple associated enzymes including pectinase, xylanase, amylase, cellulase, mannase, glucanase, glucosidase,galactosidase, feruloyl esterase, carboxypeptidase and phosphatase and is rich in natural complex enzymes is obtained by culture. Aspergillus niger is named Aspergillus niger BAK200389, and the preservation number of the Aspergillus niger is CGMCC No.19613. According to the used strain, the strain can produce multiple enzymes, and has the characteristics of stable growth, more types of produced enzymes and safety; and enzyme activity is considerable and is daily kept at 90,000-110,000, and the highest enzyme activity can reach about 120,000.

The invention discloses functional yellow wine enriched in ferulic acid and a production method of the functional yellow wine, belonging to the technical field of food brewing. The aspergillus niger CCTCC NO:M 2015202 feruloyl esterase, amylase and cellulase are high in activity, when the enzymes are added into the yellow wine fermentation process, normal fermentation of the yellow wine is not influenced, saccharifying of raw materials in the yellow wine production process is accelerated, the yellow wine production and fermentation cycle is shortened, and the original flavor and taste of the yellow wine can be maintained. The method disclosed by the invention comprises the following steps: by taking sticky rice as a raw material, inoculating aspergillus niger strain spore suspension containing high-yield feruloyl esterase during jar dropping, adding wheat bran with high ferulic acid content, performing primary fermentation, secondary fermentation, pressing, boiling wine, filling, sealing the jar and the like. The yellow wine produced by the method disclosed by the invention is enriched in ferulic acid and has the effects of resisting oxidation, reducing blood pressure, resisting thrombus, reducing blood fat, resisting bacteria, diminishing inflammation, preventing cancers, protecting the liver, treating the diabetes and the like. Moreover, the production process is energy saving and environmentally friendly, and the additional value of byproducts of wheat production is improved.

The invention discloses novel feruloyl esterase, a mutant and an application thereof, and belongs to the field of bioengineering. The method comprises the following steps: firstly, successfully screening out a feruloyl esterase gene from anaerobic fungi Neocallimatix sp. by utilizing a genomics technology; according to the preference of a prokaryotic system codon, the GC content of a feruloyl esterase gene and the secondary structure of mRNA, and codon optimization is carried out on the feruloyl esterase gene to obtain a novel feruloyl esterase gene sequence. The enzyme has an esterase regionand a glucoside hydrolase 11 region, site-specific mutagenesis is carried out on the two regions; the obtained mutant gene is also expressed in escherichia coli; separation and purification results show that the novel feruloyl esterase and the mutant thereof are successfully expressed in escherichia coli, and the esterase activity of the mutant is far lower than that of a non-mutant, so that the strain source of the feruloyl esterase is enriched, and the feruloyl esterase has theoretical guiding significance in the aspects of improving the digestibility of feed fibers and improving the production performance of animals.

The invention discloses a preparation method of feruloyl esterase enzyme-fermented feed and application thereof. The preparation method comprises the following steps: fermenting feruloyl esterase by simplified solid fermentation medium; then preparing the enzyme-fermented feed by the feruloyl esterase. The preparation method is simple; the prepared enzyme-fermented feed has improved flavor and increased palatability; moreover, the enzyme-fermented feed is safe and sanitary. Fed on the feruloyl esterase enzyme-fermented feed, the three-crossbred pig has raised feed intake and digestibility as well as obviously increased average daily gain, so that the feeding period is shortened and the economic benefit is thereby improved.

The invention belongs to the technical field of biological engineering, and particularly discloses a method for producing 4-vinyl guaiacol by fermenting bacillus circulans. The method takes the bacillus circulans as a producion strain. The method is characterized by comprising the following steps of choosing 1-2 bacillus circulans HG12 thalli from a test tube slant, inoculating to a liquid seed culture medium, and culturing to prepare an activated liquid seed; inoculating the liquid seed into a liquid fermentation culture medium according to inoculum size, and sequentially adding xylanase and feruloyl esterase, culturing to prepare fermentation liquid which contains the 4-vinyl guaiacol; carrying out filtration, extraction and standing phase splitting on the fermentation liquid to prepare extract liquid which contains the 4-vinyl guaiacol, and carrying out vacuum evaporation on the extract liquid to prepare a 4-vinyl guaiacol product. The method disclosed by the invention can realize the continuity and scale of 4-vinyl guaiacol production; the 4-vinyl guaiacol has the advantages of simplicity and convenience for production process, short production period, low production cost and great industrialized application prospect.

The invention discloses a composite cellulase system and application thereof. A method for preparing the composite cellulase system includes steps of carrying out fermentation culture on aspergillus niger with transformed feruloyl esterase genes from juniper-shaped penicillium and transformed dehydrogenase genes from to juniper-shaped penicillium obtain fermentation culture solution I; carrying out fermentation culture on trichoderma reesei to obtain fermentation culture solution II; compounding the fermentation culture solution I and the fermentation culture solution II according to a certainvolume ratio to obtain the composite cellulase system. The composite cellulase system and the application have the advantages that trichoderma reesei enzyme systems can be effectively improved by thecomposite cellulase system, and the crystalline cellulose degradation speeds of the trichoderma reesei enzyme systems further can be effectively increased; composite cellulase can be added in procedures for producing starch fuel ethanol by means of yeast synchronous saccharification and fermentation, accordingly, the viscosity of corn mash can be reduced, residual celluloses and starch further can be hydrolyzed, corn can be efficiently transformed into the fuel ethanol, and obvious economic benefits can be brought to the ethanol fermentation industry.

The method relates to a method for extracting ferulic acid esterase from rumen fluid or culture solution which produces the ferulic acid esterase, the method comprises following steps: filtering centrifugal, ultrafiltration enrichment, heating, ion-exchange chromatography, hydrophobic interaction chromatography and molecular sieve chromatography. The invention extracts various proteins which are provided with ferulic acid esterase activity from the rumen fluid or the culture solution whose components are more complex and ferulic acid esterase species are more according to different physicochemical natures of different ferulic acid esterase through organic combination of various separating and purifying steps and obtains esterase with high activity. Various ferulic acid esterase can be extracted through using the method of the invention and thereby the invention reach better production performances and has higher practical value in the feed industry.

The invention discloses a method for producing feruloyl esterase by the fermentation of aspergillus fumigatus. The activated aspergillus fumigatus with the preservation number of CGMCC No. 3.5835 is inoculated into culture medium, and is fermented under the temperature of 25 DEG C to 30 DEG C for two to six days, wherein the culture medium comprises corn starch or bran and basal liquid culture medium. The method comprises the following steps: the corn starch is adopted as the main material of the fermentation medium, the feruloyl esterase is produced after the aspergillus fumigatus is fermented in the liquid for four to five days, and the enzyme activity stably reaches more than 98U/L. The material source is wide, and the growth and multiplication of bacterial strains are fast; the technique is simple, and conditions are mild, and are easy to control; and moreover, the enzyme activity of the produced feruloyl esterase is high and highly stable, the measurement of enzyme activity is convenient, and the invention provides a new idea for the industrialized production of the feruloyl esterase.

A method for preparing low-viscosity Semen plantaginis polysaccharides through an enzyme technology comprises the following steps: 1, drying Semen plantaginis, and removing impurities; 2, extracting Semen plantaginis polysaccharides through adopting a boiling water technology; 3, purifying the Semen plantaginis polysaccharides; 4, weighing a certain amount of the purified Semen plantaginis polysaccharides, dissolving the polysaccharides according to a mass/volume ratio (mg/mL) of the polysaccharides to an aqueous solution of 1:1-5:1, adding feruloyl esterase according to an enzyme activity/mass ratio (U/mg) of feruloyl esterase to the polysaccharides of 0.1-0.5, and allowing the feruloyl esterase to act for 2-24h; reacting the obtained polysaccharide and feruloyl esterase mixed system in 40DEG C water bath; and dialyzing a solution obtained after the above reaction, concentrating after dialysis ends, and freeze-drying to obtain the low-viscosity Semen plantaginis polysaccharides. The low-viscosity Semen plantaginis polysaccharides are prepared under mild conditions; and the basic structure characteristics of the polysaccharides have no obvious change before and after enzyme technology treatment.