7 results about "Protease Gene" patented technology

The application discloses a pichia pastoris bioengineering bacterium for relieving degradation of recombinant collagen, a preparation method and application thereof. Through bioinformatics analysis, gene knockout and function verification screening, three protease genes capable of relieving degradation of collagen in a pichia pastoris host are obtained, namely, chr1-4_0611, chr1-4_0362 and chr2-2_0380. Knocking out the protease chr1-4_0611 can make the total protein yield accumulation increase to 3.5 times of that of a control bacterium, and the proportion of a target band is greater than 50%, which can provide good theoretical and technical guidance for effective expression of other recombinant proteins in the pichia pastoris host.

This invention relates to the field of biotechnology, and discloses an engineered *Yarrowia lipolytica* strain that produces p-coumaric acid using glucose as a substrate, its construction method, and its applications. Construction method: A tyrosine ammonia-lyase gene is integrated using a CRISPR / Cas9 localization and integration method. TAL In *Yarrowia lipophila* strains, the DAHP synthase gene was enhanced. ARO4 , DHS1 and AROG Overexpression of tyrosine synthase TYR Genes and histidine phosphotransferases HIS5 The protease gene was further integrated to synthesize exogenous phenylalanine deamination and hydroxylation pathway genes for p-coumaric acid, including phenylalanine ammonia-lyase gene, cinnamate hydroxylase gene, and P450 reductase gene. The p-coumaric acid produced by the engineered *Yarrowia lipolytica* strain of this invention can reach a maximum yield of 1.7 g / L in shake flasks, and a yield of 30 g / L in a 5 L fed-batch fermentation tank, demonstrating significant industrial application value.

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 present application belongs to the field of biomedical technology, and particularly relates to a method for screening of quorum sensing system inhibitors of Pseudomonas aeruginosa and simultaneously determining the target of the inhibitors. The method comprises the following steps: constructing a QS inhibitor screening system, and the QS inhibitor screening system takes protease as a screening marker; constructing a mutant with a deleted protease gene, which provides QS signal molecules in the process of co-culture; and screening a compound with inhibitory effect on the QS system and determining the target of the compound by means of co-culture or exogenous addition of QS signal molecules. The present application establishes a simple, fast, effective and high-throughput QSI screening method, and the target of the compound can be determined simultaneously.

The application belongs to the technical field of biology, and discloses application of mf1-01441 gene, protease and recombinant strain; specifically, application of mf1-01441 gene in preparation of protease and application of mf1-01441 gene in preparation of recombinant strain capable of producing protease, wherein the nucleotide sequence of the mf1-01441 gene is shown as SEQ ID NO. 1; the application successfully screens a novel protease mf1-01441 gene by constructing a Fosmid library of intestinal tract microorganisms of gibel carp; the library not only breaks through the limitation of traditional culture method, captures genetic information of a large number of uncultured microorganisms in the intestinal tract of gibel carp, lays a foundation for exploring microbial resources in the unique habitat, but also provides high-quality materials for screening of functional genes such as protease, and is expected to provide a new idea for improving feed utilization rate of fish and reducing breeding cost, and promote sustainable development of green breeding.

The present application belongs to the technical field of genetic engineering and enzyme engineering, and particularly relates to a low-temperature neutral protease from marine microorganism colwellia psychrerythraea, a preparation method thereof and application of the low-temperature neutral protease in hydrolysis of proteins. In the present application, a protease gene from colwellia psychrerythraea is connected with an expression vector pPIC9K to obtain a recombinant plasmid, then the recombinant plasmid is electrotransformed into Pichia pastoris cells, and a target gene is expressed by using methanol induction, and a centrifugal product is obtained after removing supernatant, thereby obtaining a recombinant low-temperature neutral protease. The recombinant low-temperature neutral protease can be used for hydrolysis of fish meal, and the recombinant low-temperature neutral protease has higher enzymatic activity in the range of 15 DEG C to 35 DEG C, and still maintains more than 60% at 15 DEG C, so that the recombinant low-temperature neutral protease is a neutral low-temperature metal protease which can maintain higher activity at low temperature or normal temperature.

The invention relates to the technical field of microbial genetic engineering and fermentation engineering, and particularly discloses an aspergillus oryzae genetic engineering strain and application thereof, the aspergillus oryzae genetic engineering strain is ZW28-deltasA1 deltasB1 deltasC1 deltasD1, and is a quadruple gene knockout strain constructed by taking aspergillus oryzae ZW28 as an original strain and continuously knocking out protease genes AosedA, AosedB, AosedC and AosedD by using a gene editing technology. According to the quadruple gene knockout strain provided by the invention, the extracellular total protease activity is averagely reduced by 35%, the yields of expressed human lysozyme and calf chymosin respectively reach 65 mg / L and 122.2 mg / L, which are 4.7-5.2 times higher than those of an original strain, and the quadruple gene knockout strain shows strong universality for foreign proteins with different sources and pH characteristics, and provides a dominant platform for efficient production of industrial enzymes.