26 results about "Pectin lyase" patented technology

The invention discloses a lactobacillus lactis for relieving constipation and a preparation method thereof, target strains are respectively separated from a fermented bean curd sample and screened by a pectinase screening agar culture medium, and are named as lactobacillus plantarum PD-14 and lactobacillus pentosus PD-16, the lactobacillus lactis has a gene for synthesizing pectate lyase, and the lactobacillus lactis has a gene for synthesizing pectate lyase. Relates to the microbial field. The invention provides a beneficial bacterium for relieving constipation and a preparation method thereof, wherein the beneficial bacterium is capable of efficiently relieving constipation, low in cost, stable and easy to industrialize.

This invention discloses a nicotine-resistant pectin lyase mutant and its application in tobacco leaf treatment. Using pectin lyase PL6 as the starting enzyme, this invention designs and optimizes mutation sites, designing single-site saturation mutations based on the two hydrogen bond formation sites between the enzyme and nicotine, Lys-107 and Asp-84, to construct mutants. The pectin lyase mutants D84A and K107R of this invention retain 83.63% and 81.30% of their enzyme activities at a 1% nicotine concentration, respectively. Spraying tobacco leaves with D84A and K107R enzyme solutions significantly improves tobacco leaf quality due to pectin degradation.

A process for producing three recombinant enzymes capable of hydrolyzing each class of organophosphate, carbamate, and pyrethroid insecticides, as well as a formulation containing these enzymes provides the recombinant plasmid DNA vector, in which the protein secretory system has been modified to enhance the secretion of recombinant protein into the periplasmic space or culture medium. A nucleotide sequence encoding PelB (pectate lyase B) signal sequence for periplasmic localization is deleted and replaced with a nucleotide sequence encoding LamB (maltoporin or phage lambda receptor) signal sequence to enable a more efficient secretory recombinant protein. Three recombinant plasmid DNA vectors for the expression of recombinant proteins that are constructed using the modified plasmid DNA vector described above. The recombinant proteins include a recombinant organophosphate-hydrolyzing enzyme; a recombinant carbamate-hydrolyzing enzyme; and a recombinant pyrethroid-hydrolyzing enzyme.

The invention discloses a heterologous expression pectate lyase engineering bacterium, a method for preparing pectate lyase by using the heterologous expression pectate lyase engineering bacterium and application of the heterologous expression pectate lyase engineering bacterium. Heterologous expression pectate lyase is recombinant protein optimized by codons, and the amino acid sequence of the heterologous expression pectate lyase is shown as SEQ ID NO.2; and the nucleotide sequence of the coding gene is as shown in SEQ ID NO.3. According to the invention, a recombinant expression vector containing the coding gene is constructed, pichia pastoris X-33 is used as host bacteria to construct engineering bacteria, and efficient expression of the pectate lyase is realized by optimizing vector elements and fermentation conditions. And the production time consumption is short, the cost is low, the process is easy to control, and the method is suitable for industrialization. And the produced enzyme has good purity, activity and stability, can efficiently degrade pectin substances when being used for tobacco, and improves the sensory quality and safety of cigarettes.

The invention discloses a mixed fermentation brewing method of Xuangxiang kiwi fruit and sunshine rose grape composite fruit wine, and belongs to the technical field of fruit wine brewing. According to the method, Xuangxiang kiwi fruits and sunshine rose grapes are used as raw materials and are compounded according to the mass ratio of 2: 1, SO2 and pectin lyase are added, independently-bred pichia kluyveri HN22 and commercial saccharomyces cerevisiae RV171 are adopted for mixed fermentation, low-temperature natural clarification, cold stabilization, multi-stage membrane filtration and other treatment are performed, and the high-purity kiwi fruits and sunshine rose grapes are obtained. The composite fruit wine is rich in fragrance, moderate in acidity and low in methanol content. According to the preparation method, raw material ratio optimization is combined with a mixed fermentation process, so that the problems that the traditional kiwi fruit wine is boring in aroma and sour and astringent in taste and methanol is easy to exceed the standard are solved, the sensory quality of a product is remarkably improved, and the preparation method has a good industrialization prospect.

The invention provides a transcription factor complex for regulating and controlling the softening process of strawberry fruits and application of the transcription factor complex, and relates to the technical field of agricultural biology. The complex is composed of FanMYB10. B1, FanbHLH1. B1 and FanWD40. D1, and the nucleotide sequences of the FanMYB10. B1, the FanbHLH1. B1 and the FanWD40. D1 are respectively as shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3. The defects in the prior art are overcome, the complex regulates and controls the fruit softening by activating the transcription efficiency of a pectate lyase gene PL promoter, meanwhile, it is verified that FanMYB10.B1 expression can improve the quality of strawberry fruits after picking, an abscisic acid-MBW transcription complex-pectate lyase cascade regulation and control network is disclosed, and in order to delay the softening of the strawberry fruits, the abscisic acid-MBW transcription complex-pectate lyase cascade regulation and control network is provided. A target spot is provided for improving the postharvest quality of fruits.

The invention provides the method for fluorescence quantitative determination of the activity of the endo-pectinase, the enzyme activity of the endo-pectinase can be accurately detected by constructing a proper endo-pectinase enzyme activity detection system, the method is green, safe, convenient to operate and low in cost, the defects of the existing detection method can be avoided, and the method can be widely applied to detection of the endo-pectinase activity of the endo-pectinase activity of the endo-pectinase activity of the endo-pectinase activity of the endo-pectinase activity of the endo-pectinase. The application prospect is wide.

The invention discloses a preparation method of a flaxseed extract and an antioxidant and anti-inflammatory composition based on the flaxseed extract, and belongs to the technical field of natural product extraction and medical products. The preparation method comprises the following steps: firstly, improving the anti-oxidation effect of the flaxseed extract by improving the preparation mode of the flaxseed extract, particularly performing two-step specific biological enzymolysis: performing first-step enzymolysis by adopting pectin lyase shown as SEQ ID NO.1 and rhamnogalacturonan lyase shown as SEQ ID NO.2; cellulase, xylanase and arabinofuranosidase are adopted for second-step enzymolysis, and the DPPH free radical scavenging rate of the obtained product is higher than that of a traditional extraction method. On the basis, the flaxseed extract is compounded with tetrahydrocurcumin and dihydroquercetin, the ratio is further optimized, the antioxidant and anti-inflammatory composition is obtained, and the antioxidant and anti-inflammatory effects of the composition are remarkably superior to those of a single component.

The application discloses a alginate lyase mutant E87Q, a coding gene, a recombinant expression vector and a genetically engineered bacterium, and belongs to the technical field of genetic engineering. The application takes alginate lyase PpAly7A as a starting template, and a mutant with improved relative specific activity is obtained through site-directed mutagenesis. Compared with the wild enzyme PpAly7A, the mutant E87Q has a relative specific activity increased to 133.82%, and the corresponding site can be used as a new target for subsequent rational design.

The invention belongs to the technical field of food, and more particularly relates to grape wine and a brewing method thereof, and the brewing method specifically comprises the following steps: after high-pressure pulse treatment of grape pulp, adding pectin lyase and grape seed polyphenol extract, and carrying out cold dipping and enzymolysis for 60-72 hours at 5-8 DEG C under the protection of inert gas to obtain enzymatic hydrolysate; inoculating a composite yeast agent into the enzymatic hydrolysate, and performing staged temperature-controlled fermentation; and after the fermentation is finished, adding nanoscale oak microcapsules into the fermentation liquor, and ageing for 3-4 months in a micro-aerobic environment with dissolved oxygen of 0.2-0.5 mg / L to obtain the grape wine. According to the method, the polyphenol extraction rate of the wine is increased to 92%, esters are larger than or equal to 200 mg / L, melatonin is 0.5-2 mg / L, methanol is smaller than or equal to 80 mg / L, acetaldehyde is smaller than or equal to 20 mg / L, and the ageing period is shortened to 3 months.

The invention discloses a preparation method of a jaboticaba pomace extract with an anti-oxidation function, and the jaboticaba pomace extract is prepared by taking jaboticaba pomace as a raw material and adopting a compound extraction process of coupling pectin lyase enzymolysis with physical wall breaking homogenization. Meanwhile, the invention further discloses a jaboticaba pomace extract with an antioxidant function, the anthocyanin extraction rate of the jaboticaba pomace extract is 7.9-8.6 mg / g, and the polyphenol extraction rate of the jaboticaba pomace extract is 60-66 mg / g. According to the jaboticaba pomace extract with the anti-oxidation function and the preparation method of the jaboticaba pomace extract, the extraction efficiency and the yield of polyphenol and anthocyanin in pomace are greatly improved by aiming at a compound extraction process for synergistically and efficiently extracting polyphenol and anthocyanin from jaboticaba pomace through enzymolysis coupling with physical wall-breaking homogenization of pectate lyase; the antioxidant performance of the extract is obviously superior to that of a commercially available common antioxidant vitamin C, under the condition of the same concentration, the scavenging rates of the extract on DPPH and ABTSfree radicals are more than 5 times that of the vitamin C, and the extract has outstanding activity.

The invention belongs to the technical field of plant molecular biology, and discloses a soybean disease resistance-related protease GmRD21A gene, a coding protein thereof, and an application of the gene in plant disease resistance. The nucleotide sequence of the protease GmRD21A gene is as shown in SEQ ID NO. 1, and the amino acid sequence of the polypeptide coded by the protease GmRD21A gene is as shown in SEQ ID NO. 2. Protein spectrum analysis of phytophthora sojae pectate lyase PsPL1 proves that the protease GmRD21A degrades pectate lyase secreted by phytophthora and can resist attack of pathogenic bacteria pectate lyase on plant cell walls, and meanwhile, overexpression of the GmRD21A gene in soybeans can remarkably improve the resistance of plants to phytophthora. The invention can be applied to crop breeding anti-epidemic disease improvement, and is hopeful to improve the disease resistance of plants to epidemic diseases, thereby achieving the purposes of increasing yield and reducing pesticide consumption.

The application discloses a brown algae pectin lyase and application thereof in preparation of guluronate oligosaccharide, and belongs to the technical field of biotechnology, and particularly relates to a brown algae pectin lyase and application thereof in preparation of guluronate oligosaccharide. The protein of the application is any one of the following: 1) a protein with the amino acid sequence of sequence 1 in the sequence list; 2) a protein with the amino acid sequence of the 22th to 453th of sequence 1 in the sequence list. The application purifies a recombinant brown algae pectin lyase, and the specific enzyme activity of the purified enzyme is 86.1 U / mg, and the recovery rate is 76%. The optimal pH of the brown algae pectin lyase VaAly7 is 7.0, and the brown algae pectin lyase is stable between pH 4.0 and 9.5; the optimal temperature is 30 DEG C, and the brown algae pectin lyase is stable below 30 DEG C, and is a cold-adapted brown algae pectin lyase, and has the advantages of easy control and energy saving in production and application, and has application potential in production of guluronate oligosaccharide.

The invention discloses a persimmon macromolecular polyphenol extract as well as a preparation method and application thereof, and relates to the technical field of plant active ingredient extraction. In the enzymolysis process, only pectin lyase (EC 4.2. 2.10) is used for enzymolysis, and the method specifically comprises the following steps: step 1, pulping a persimmon raw material to obtain persimmon pulp; step 2, adding pectin lyase into the persimmon pulp obtained in the step 1, and carrying out enzymolysis extraction; and step 3, centrifugally filtering the enzymolyzed persimmon pulp obtained in the step 1 to obtain an extracting solution, and carrying out ultrafiltration, concentration and freeze drying on the extracting solution to obtain the extract. The invention discovers and verifies that the pectate lyase has unexpected high efficiency and specificity for degrading green persimmon cell wall pectin and releasing bound polyphenol at normal pressure for the first time, and the effect of the pectate lyase is far superior to that of common pectinase and cellulase. The obtained extract has extremely high antioxidant activity, the DPPH free radical scavenging rate is larger than 91%, and it is indicated that the extract has very high biological utilization value and development potential.

The invention provides a transcription factor FanPRE-1. B1 for regulating and controlling softening of strawberry fruits and application of the transcription factor FanPRE-1. B1, and relates to the technical field of agricultural biology. The nucleotide sequence of the transcription factor FanPRE1. B1 is as shown in SEQ ID NO: 1, and the quality of picked fruits can be improved by inhibiting the expression of the FanPRE1. B1; a downstream gene regulated and controlled by FanPRE1.B1 is identified, and the downstream gene is a pectin lyase gene; the FanPL7a1. A1, the FanPL7a1. A1, the FanPL7a1. B1, the FanPL7b1. A1, the FanPL7b1. B1, the FanPL7b1. C1 and the FanPL7b1. D1 are specifically included, and the FanPL7a1. A1, the FanPL7b1. B1, the FanPL7b1. C1, the FanPL7b1. D1 and the The expression level of any one of the genes can be reduced by inhibiting the expression of FanPRE1.B1, so that the degradation of pectin is inhibited, and the softening of fruits is delayed. The invention provides a new thought for delaying softening of strawberry fruits and improving the quality of picked fruits.

The invention provides a method of producing a hydrolysed pectic polysaccharide isolate that is enriched in rhamnogalacturonan-I, said method comprising the steps of:providing a pectin-rich substrate that has been obtained from plant material without the use of organic solvent, said pectin-rich substrate containing at least 3% by weight of dry matter of pectic polysaccharides;subjecting the pectin-rich substrate to enzymatic treatment to partially hydrolyse the pectic polysaccharides, said treatment enzymatic treatment comprising the use of one or more pectinases selected from pectin lyase (EC4.2.2.10), pectate lyase (EC 4.2.2.2), rhamnogalacturonan galacturonohydrolase (EC 3.2.1.173), endo-polygalacturonase (EC 3.2.1.15), exopolygalacturonase (EC 3.2.1.67 and EC 3.2.1.82);subjecting the partially hydrolysed pectic polysaccharides to ultrafiltration using an ultrafiltration membrane having a molecular weight cut-off in the range of 5 to 100 kDa; andrecovering the ultrafiltration retentate.The present invention further relates to the hydrolysed pectic polysaccharide isolate obtained by the present method and to a process of preparing a product selected from a nutritional formulation, a food product, a dietary supplement, a beverage or a pharmaceutical product, said process comprising addition of the aforementioned hydrolysed pectic polysaccharide isolate.

The invention belongs to the field of development and utilization of beautiful millettia roots, and particularly relates to a beautiful millettia root flower extract and a preparation method and application thereof. Fresh flower buds in the initial flowering period of beautiful millettia roots are adopted as raw materials, fermentation liquor is prepared through fermentation of lactobacillus, lactobacillus casei and lactobacillus acidophilus, after sterilization, cellulase, pectin lyase and cellobiohydrolase are added for enzymolysis, enzymatic hydrolysate is obtained, after enzyme deactivation of the enzymatic hydrolysate, alcohol precipitation is conducted to remove precipitates, and the finished product is obtained. The prepared millettia specisoa flower extract can reduce blood sugar of rats with type 2 diabetes mellitus and regulate intestinal flora of the rats, and a new candidate medicine is provided for treatment of diabetes mellitus.

The application discloses a brown alga gelolysis enzyme mutant E180Q, a coding gene, a recombination expression carrier and a gene engineering bacteria, and belongs to the technical field of gene engineering. The application takes brown alga gelolysis enzyme PpAly7A as a starting template, and obtains a mutant with improved thermal stability through site-directed mutation and combined mutation. The thermal stability is improved by destroying the salt bond of Glu180-Arg178 (E180Q), the half-life of the mutant at 50 DEG C is improved to 7.246 days compared with wild enzyme PpAly7A, and the site can be used as a new target for subsequent rational design.

The invention discloses high-temperature alkaline pectin lyase as well as a gene and application thereof. The high-temperature alkaline pectin lyase is derived from pectin lyase MtPLY-1 and MtPLY-2 from the mycelia thermophila. The invention also discloses a method for preparing the pectin lyase MtPLY-1 and the pectin lyase MtPLY-2 through high expression and production of the destructor thermophilus, and an application of the pectin lyase MtPLY-1 and the pectin lyase MtPLY-2 in degradation of tobacco biomass as well as a method for characterizing enzyme properties of the pectin lyase MtPLY-1 and the pectin lyase MtPLY-2 and the application of the pectin lyase MtPLY-1 and the pectin lyase MtPLY-2 in degradation of tobacco biomass. The alkaline pectin lyase provided by the invention is good in thermal stability and wide in temperature application range, shows relatively good enzyme activity at the temperature of 5070 DEG C and the pH value of 7.010.0, shows a good synergistic effect on cellulase, can effectively promote degradation and utilization of tobacco waste biomass and improve the sensory quality of tobacco leaves, and thus, has relatively high application value.

A highly active mutant pectin lyase and its applications are disclosed, specifically mutant pectin lyases PGLA-rep4, PGLA-rep1, and PGLA-rep2. The modified pectin lyases of this invention exhibit enhanced enzyme activity and have broad application prospects in pulping, papermaking, textiles, feed, and other fields.

The invention discloses application of combination of pectin methylesterase and pectin lyase in degradation of tobacco leaf pectin, and is characterized in that the application comprises the following steps: weighing recombinant pectin methylesterase according to a proportion of 0.01-0.05% of the mass of tobacco leaves, weighing recombinant pectin lyase according to a proportion of 0.01-0.05% of the mass of the tobacco leaves, respectively diluting with deionized water, and mixing to obtain an enzyme mixed solution; uniformly spraying the enzyme mixed solution on the surfaces of the tobacco leaves; the sprayed tobacco leaves are subjected to fermentation treatment; after fermentation treatment is completed, heating the tobacco leaves to inactivate enzyme; wherein the nucleotide sequence of the recombinant pectin methylesterase is as shown in SEQ ID NO.3; and the nucleotide sequence of the recombinant pectin lyase is as shown in SEQ ID NO.6. The high-purity active enzyme is obtained through pichia pastoris heterologous expression, and the method is suitable for large-scale production; through the synergistic effect of the two enzymes, the pectin degradation rate of the tobacco leaves reaches 35.2%, the total sensory quality is improved by 2.68, and the degradation rate is higher than that of a single enzyme; the application amounts of the two enzymes are 0.03% respectively, namely the optimal effect is achieved, and the cost is controlled.

The invention belongs to the field of biological enzyme preparations, and particularly relates to a compound enzyme preparation for improving combustibility of Nanping cigars and application of the compound enzyme preparation. According to the method, the Nanping cigar tobacco leaves with improved combustion performance are prepared by harvesting the Nanping cigars, airing the Nanping cigars, screening and compounding enzyme preparations, optimizing an enzymolysis mode and controlling enzymolysis parameters only by adopting a compound enzyme preparation without non-enzyme preparations. Pectate lyase and pectin esterase in pectinase, papain in protease, beta-amylase saccharifying enzyme and isoamylase in amylase and cellulase are screened out and combined, and further optimized that pectate lyase, pectate esterase and cellulase are subjected to mixed enzymolysis, enzymolysis is carried out, enzymolysis is carried out, and enzymolysis is carried out. The papain, beta-amylase and isoamylase solutions are mixed for enzymolysis, and the temperature and time of two times of enzymolysis are controlled, so that the combustibility of the Nanping cigar tobacco leaves is remarkably improved, and meanwhile, the smoking evaluation result of the Nanping cigar tobacco leaves is good.

This invention relates to the field of biotechnology, and more particularly to the tobacco pectin lyase NtPL gene and its applications. This invention is the first to experimentally demonstrate that mutants lacking pectin lyase function (as shown in SEQ ID NO:2) exhibit reduced petiole stress tolerance and increased leaf drop, while overexpressed mutants show increased petiole stress tolerance and reduced leaf drop. This provides a new means to regulate the mechanical strength of tobacco petioles and offers a new strategy for tobacco molecular breeding, which is of great significance for stabilizing tobacco yield and quality.

The invention discloses radix ophiopogonis powder capable of improving mitochondrial activity and resisting aging and a preparation method and application of the radix ophiopogonis powder, and relates to the technical field of radix ophiopogonis products. The prepared radix ophiopogonis powder can effectively improve mitochondrial membrane potential, activate a mitochondrial compound I, increase ATP (adenosine triphosphate) generation and reduce ROS (reactive oxygen species) level, remarkably recover heart beat output and realize cross-model mitochondrial activation in a zebra fish myocardial injury model, and can remarkably improve activity of a nematode compound I, nematode movement ability and nematode life in a nematode model. Meanwhile, the compound has good safety in a human body, and can improve mitochondrial activity and delay telomere shortening after being orally taken. The composition is used for human skin, has excellent moisturizing, barrier repairing and anti-oxidation effects, and has the functions of remarkably improving mitochondrial activity and resisting aging.

The application discloses a pectin lyase mutant ADdPelZ, a coding gene thereof, a preparation method and application, wherein the pectin lyase mutant ADdPelZ is obtained by mutating an amino acid at the 168th position of a substrate binding pocket of a wild-type pectin lyase DdPelZ from a macromolecular branched valine into a positively charged arginine, and mutating an amino acid at the 319th position of a rigid region from a macromolecular branched valine into a small-molecular-weight alanine; the amino acid sequence and the nucleotide sequence of the mutant are respectively represented as SEQ ID NO. 1 and SEQ ID NO. 2. The mutant enzyme provided by the application has obvious improvements in enzyme activity and heat resistance under alkaline conditions, and solves the problems of low catalytic activity and insufficient heat resistance of the wild-type pectin lyase under alkaline conditions, thereby creating good conditions for application of the enzyme in cotton and hemp processing, pulping and papermaking, and feed processing.

The invention provides modified dietary fiber with satiety and a preparation method of the modified dietary fiber. The preparation method comprises the following steps: soaking plant raw materials at 4 DEG C, performing liquid nitrogen freezing and crushing to 30-50 microns, and performing vacuum drying to obtain superfine powder; carrying out step-by-step enzymolysis on the superfine powder through four enzymes (alpha-amylase, alkaline protease, pectin lyase and cellulase) to obtain enzymatic hydrolysate; adding degradable starch microspheres, stirring, standing at low temperature, centrifugally washing, freezing, and removing the microspheres through saccharifying enzyme to obtain hierarchical porous dietary fibers; after urea-glycerol regulation and high-speed shearing, gradient pulse microjet treatment, low-temperature heat preservation and drying are adopted, and a finished product is obtained. The modified dietary fiber with a high specific surface area and a multi-stage through pore channel structure is constructed through a pore forming technology and gradient pulse microjet, a foundation is laid for an efficient satiety function from the physical structure level, and the technical problems that traditional dietary fibers are single in pore channel and insufficient in adsorption and expansion performance are solved.