Neutral phytase transgenic recombinant lactobacillus and product and application thereof

Abstract

The invention relates to a neutral phytase transgenic recombinant lactobacillus and a product and application thereof. Neutral phytase genes in AY220075 Bacillus subtilis with lactobacillus signal peptides are integrated in a genome, and an amplified sequence is a 1-1152bp basic group; signal peptide genes are 260-349th site basic groups of Z14250; and the neutral phytase genes and the signal peptide genes are inserted into multiple cloning sites after being connected and are integrated into ribosome binding sites of Lactobacillus casei L.CECT5276. The neutral phytase transgenic recombinant lactobacillus provided by the invention can be used as a feed additive for pig husbandry, can make up the defect that the intestinal tract of a pig cannot secrete phytase, can promote the metabolism of phytate phosphorus in a feedstuff raw material and reduce discharge of inorganic phosphorus, which are favorable for the protection of the environment. A host bacteria lactobacillus also has beneficial function to animal bodies, and has important application value in production of pig, poultry and other monogastric animals.

Description

technical field [0001] The invention relates to a gene recombination technology, in particular to a neutral phytase gene-transferred lactobacillus and its product and application. Background technique [0002] Phytase (Phytase) is myo-inositol hexaphosphate phosphor-hydrotase, which catalyzes the hydrolysis of phytic acid (ie, phytic acid) and phytate into inositol and phosphoric acid (or phosphate). A general term for a class of enzymes. The chemical name for phytic acid is cyclohexyl hexaphosphate. The molecular formula is C6H18O24P6, and the structural formula is C6H6 [OPO(OH) 2 ] 6 , it is a light yellow or tan viscous liquid. Soluble in water, ethanol, acetone, almost insoluble in benzene, chloroform, ether and hexane. Although phytic acid itself has little toxicity, it has strong chelating ability, so it has anti-nutritional effect. It will interact with a variety of metal ions such as Zn in the gastrointestinal tract of animals 2+ , Ca 2+ 、Cu 2+ , Fe 2+ Pla...

AI Technical Summary

Problems solved by technology

In the seeds of grains, beans, and oil crops, the basic storage form of phosphorus is phytate phosphorus, which accounts for 60%-80% of the total phosphorus in plants[3], but it is difficult to be utilized by monogastric animals, resulting in great Problem: First is the waste of phosphorus resources (Ehrlich KC, Montalbano B G, Mullaney E J, et al. Identification and cloning of a second phytase gene (phyB) from Aspergillus niger (ficuum) . Biochemical and Biophysical Research Communications, 1993, 195:53 ~57.)

[0011] At present, acid phytase has been commercialized, but there are still some problems: first, the unit enzyme activity of feed products is still relatively low; second, there are few studies on neutral phytase, and no corresponding products have been put on the market; Enzyme Product Thermal Stability Issues

At present, there have been many successful reports on the transformation of the thermal stability and catalytic pH of acid phytase by using genetic engineering and protein engineering technology, but there is no similar report on neutral phytase.

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