8 results about "Feruloyl esterase" patented technology

Provided are the screening and identification of a Pseudomonas sp. strain for producing feruloyl esterase at high yield and a use of the Pseudomonas sp. strain as a bioaugmentation agent in fermented foods, relating to the technical field of brewing. By means of screening, a Pseudomonas sp. strain A-4H for producing feruloyl esterase at high yield is obtained. After cultivation for 36 h, the Pseudomonas sp. strain A-4H can produce ferulic acid at a concentration of 153.79±2.36 mg / L, and the activity of feruloyl esterase in a fermentation broth reaches 136.63±10.27 U / L. The addition of the Pseudomonas sp. A-4H during the brewing of Huangjiu can effectively increase the content of ferulic acid, the content of phenolic acid, and the content of organic acid in Huangjiu, providing an effective biological pathway for increasing the content of ferulic acid in fermented foods.

The application discloses a recombinant feruloyl esterase and a mutant and application thereof, and belongs to the technical field of enzyme engineering. Clostridium butyricum, The wild-type enzyme amino acid sequence is shown as SEQ ID NO:1, and the coding gene sequence is shown as SEQ ID NO:2; the target enzyme is obtained by constructing a recombinant expression vector containing the CbFAE gene, inducing expression in Escherichia coli BL21 (DE3) and purifying; substrate spectrum analysis shows that the enzyme not only has catalytic decomposition activity of model substrates such as methyl ferulate, but also has catalytic activity of decomposing mono (2-hydroxyethyl) terephthalate (MHET) into terephthalic acid (TPA) and ethylene glycol (EG); the application also constructs the CbFAE mutant L168R based on sequence and structure analysis, the amino acid sequence of the mutant enzyme is shown as SEQ ID NO:3, and the catalytic MHET decomposition activity of the mutant enzyme is increased by nearly 4 times compared with the wild type under the same reaction condition. The recombinant feruloyl esterase and the mutant provided by the application expand the types of MHET hydrolytic enzymes and provide a new way for the biodegradation of MHET.

The invention belongs to the technical field of genetic engineering and biology, and particularly discloses construction and application of a xylan feruloyl esterase delivery plasmid. The amino acid sequence of the xylan feruloyl esterase is shown as SEQ ID NO: 1, a xylan feruloyl esterase delivery plasmid pQ-XFE based on IncQ type plasmid and a recombinant genetic engineering bacterium are constructed, and thus the xylan feruloyl esterase is obtained. According to the delivery plasmid, efficient and stable delivery of xylan feruloyl esterase genes in various microorganisms is achieved, the enzyme expression quantity and activity can be remarkably improved, and the application range of the xylan feruloyl esterase genes is widened. The recombinant xylan feruloyl esterase has a remarkable degradation effect on ester compounds and can be applied to the fields of food, pharmacy, papermaking, feed and the like due to the specific activity and enzymatic characteristics of the recombinant xylan feruloyl esterase.

The application provides ferulic acid esterase mutants with improved specific activity and application thereof, and the application carries out directional evolution on a ferulic acid esterase gene derived from Bacillus pumilus, expresses the ferulic acid esterase gene in an E. coli expression system, and obtains ferulic acid esterase mutants DV1 and DV2 with improved enzyme activity and expression efficiency through screening. The amino acid sequences of the ferulic acid esterase mutants DV1 and DV2 are shown in SEQ ID NO. 4 and SEQ ID NO. 6 respectively, and the mutants DV1 and DV2 obtained by the application have higher protein yield, higher specific activity of methyl ferulate esterase, and higher degradation efficiency of macromolecular p-nitrophenol ester substrates compared with the original bacteria BpFAE. Therefore, the mutants can be used for degrading plant fibers and preparing feed fermenting agents, and have good market application prospect.

The invention provides feruloyl esterase, a DNA molecule for coding the feruloyl esterase, a recombinant expression vector, a genetically engineered bacterium and application of the feruloyl esterase in production of the feruloyl esterase, and belongs to the technical field of microbial genetic engineering. The nucleotide sequence of the DNA molecule for coding the feruloyl esterase is as shown in SEQ ID No. 1, and the amino acid sequence correspondingly coded by the DNA molecule is as shown in SEQ ID No. 2. The invention also discloses a recombinant expression vector and a genetically engineered bacterium which comprise the DNA molecule for coding the feruloyl esterase. The feruloyl esterase BgFae2148 is subjected to genetic engineering modification to obtain a corresponding expression vector and an engineering bacterium, and the produced feruloyl esterase has relatively high enzymatic activity and yield; by optimizing the selection of carriers and the optimization of induction conditions, large-scale fermentation and industrial production are realized, ferulic acid is released through enzymolysis, high-value utilization of wastes such as straws is promoted, and pollution is reduced.

A method of treating a sunflower plant stem is disclosed. The method comprises the steps of:selecting a sunflower plant, removing the flower head and leaves from the stem and preparing the stem for enzymatic treatment by slitting the stem lengthways, removing the inner core from the stem;a) treating the cut stem with a pectinase containing a first buffer solution, for 24 hours with shaking at a pH of 3.8-8.5, and at a temperature of from 20-60° C.;b) removing the stem and washing with water,c) placing the stems into a second buffer solution containing xylanase, at a pH of from 7.0-8.0 and at a temperature of from 37-50° C. for 24 hours with shaking;d) removing the stems and washing with watere) placing the stems into a third buffer solution containing feruloyl esterase, at a pH of 7.2-8.0 containing 0.01-0.05% feruloyl esterase, at a temperature of from 37-50° C. for up to 24 hours,f) removing the stems and washing with water,g) pressing the stems to remove unwanted plant matter.

The invention discloses recombinant feruloyl esterase as well as a mutant and application thereof, and belongs to the technical field of enzyme engineering. The feruloyl esterase CbFAE is derived from Clostridium butyricum, the amino acid sequence of a wild enzyme is as shown in SEQ ID NO: 1, the sequence of an encoding gene of the feruloyl esterase CbFAE is as shown in SEQ ID NO: 2, and a target enzyme is obtained by constructing a recombinant expression vector containing a CbFAE gene and performing induced expression and purification in escherichia coli BL21 (DE3). Substrate spectrum analysis shows that the enzyme not only has the activity of catalyzing decomposition of mode substrates such as methyl ferulate, but also has the catalytic activity of decomposing mono (2-hydroxyethyl) terephthalate (MHET) into terephthalic acid (TPA) and ethylene glycol (EG); the invention also constructs a CbFAE mutant L168R based on sequence and structure analysis, the amino acid sequence of the mutant enzyme is as shown in SEQ ID NO: 3, and the activity of catalyzing MHET decomposition under the same reaction condition is nearly 4 times higher than that of a wild type. According to the recombinant feruloyl esterase and the mutant thereof provided by the invention, the variety of MHET hydrolase is expanded, and a new way is provided for biodegradation of MHET.

The invention relates to lactobacillus pentosus AR10 for producing feruloyl esterase and application of the lactobacillus pentosus AR10. The lactobacillus pentosus AR1 disclosed by the invention is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number of the lactobacillus pentosus AR1 is CCTCC NO: M 20251084. The lactobacillus pentosus AR1 disclosed by the invention has the advantages that the lactobacillus pentosus AR1 is preserved in the China Center for Type Culture Collection (CCTCC); the strain provided by the invention can produce feruloyl esterase to degrade ferulate, also shows relatively strong environmental adaptability, and can still have certain growth activity in an environment with the pH value of 3.0, the temperature of 45 DEG C and 10% NaCl. In addition, the strain can also utilize various sugar source substances. When the strain is compounded with fermentation bacteria such as lactobacillus plantarum and the like to be applied to corn straw silage, the fiber content in straw can be remarkably reduced, and the nutritional value of silage is improved.