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Trans-cellulase gene recombinant lactobacilli and product and application of trans-cellulase gene recombinant lactobacilli

A technology of lactobacillus and cellulase, which is applied in application, genetic engineering, plant gene improvement, etc., can solve the problems of enzyme activity reduction and inactivation, and achieve the effects of improving digestion and absorption, reducing feed cost, and improving production performance

Inactive Publication Date: 2012-09-19
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although there are mature products of cellulase on the market, these enzyme products can only be introduced to the market after a series of processes such as microbial fermentation, separation, purification, and mixing with corresponding carriers, and these enzymes are still in storage and application. Sometimes there are strict conditions, such as temperature, pH, etc., and it is easy to be affected by external conditions to reduce the enzyme activity or even inactivate it.

Method used

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  • Trans-cellulase gene recombinant lactobacilli and product and application of trans-cellulase gene recombinant lactobacilli
  • Trans-cellulase gene recombinant lactobacilli and product and application of trans-cellulase gene recombinant lactobacilli
  • Trans-cellulase gene recombinant lactobacilli and product and application of trans-cellulase gene recombinant lactobacilli

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Effect test

Embodiment 1

[0160] Example 1: Construction of recombinant lactobacillus that secretes and expresses cellulase

[0161] 1. Cloning of cellulase gene

[0162] Using the Bacillus sp.5H genome as a template, using P1 and P2 as primers to amplify the cel gene (see attached figure 1 ), there is a specific band at about 1500bp, the purified product is connected with pMD18-T, and the plasmid pMD-cel is obtained, and its PCR identification results are shown in the attached figure 2 (A specific band can be obtained at 1500bp).

[0163] 2. Cloning of the signal peptide sp gene of Lactobacillus pumilus

[0164] Lactobacillus pumilus L.brevis1.2028 genome was used as template, and sp gene was amplified with P3 and P4 as primers (see attached image 3 ), there is a specific band at about 90bp, the purified product is ligated with pMD18-T to obtain the plasmid pMD-sp, the results of enzyme digestion and PCR identification are shown in the attached Figure 4 (PstI / XbaI double digestion can produce ...

Embodiment 2

[0169] Embodiment 2: Experiment of the influence of recombinant lactobacillus of the present invention on pig production performance

[0170] 1. Selection, grouping and feeding management of experimental animals

[0171] The experiment was carried out in the same double-row closed piggery, and all the methods of free feeding and drinking were adopted.

[0172] Fifty-four Du×Chang×Da Sanyuan hybrid growth pigs of about 20 kg were selected, with half male and half female, and they were castrated and epidemic-prevented according to routine. The initial weight was recorded, and they were randomly divided into 3 groups, with 3 repetitions in each group and 6 pigs in each repetition. Each group is processed as follows respectively: I group (control group), basal diet+0.10% secondary meal; II group (non-recombinant lactobacillus group): the non-recombinant lactobacillus preparation of basal diet+0.10% (0.10% secondary powder+ 10 8 CFU / g non-recombinant lactobacillus); III group (r...

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Abstract

The invention relates to trans-cellulase gene recombinant lactobacilli and a product and application of the trans-cellulase gene recombinant lactobacilli. Protein coded by the trans-cellulase gene recombinant lactobacilli has an amino acid sequence shown in a sequence table SEQ ID NO.1. The trans-cellulase gene recombinant lactobacilli built by the invention can be used as an oral feed additive applied to the pig industry or poultry farming so as to compensate the defects that intestinal tracts of pigs, and birds cannot secrete cellulose without adding expensive cellulase preparation, not only facilitates metabolism of crude fiber in feed material, but also can promote growth of pigs and increase the economic benefits of cultivation. In addition, the host bacterium lactobacillus is lactobacillus casei, and has a probiotic function to an animal body. The invention provides an artificial cellulase probiotic genetic engineering lactobacillus, which has important application value in production of monogastric animals such as pigs, birds and the like.

Description

technical field [0001] The invention relates to a cellulase gene-transformed lactobacillus, and also relates to an application of the cellulase gene-transformed lactobacillus as a feed additive in feed. Background technique [0002] Cellulase is a general term for a group of enzymes that degrade cellulose, mainly composed of exo-β-glucanase, endo-β-glucanase and β-glucosidase. Due to the huge market potential of cellulase in the fields of feed, alcohol, textile and food, it has been favored by people in the industry at home and abroad. It will be the fourth largest industrial enzyme after glucoamylase, amylase and protease, even in China. It is entirely possible to become the largest enzyme species [YANG Sheng, HOU Hong-ping, et al.Recent advance in cellulase[J]. A new growth point in . [0003] Cellulase is widely used. In terms of feed, for ruminants, forage cellulase can supplement the deficiency of cellulase formed by microorganisms in the body, promote the growth of b...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/74C12N15/31C12N15/56C12N9/42A23K1/16C12R1/225
Inventor 丁轲余祖华程相朝闫文朝刘炜刘一尘胡益源黄俊克王天奇赵振升
Owner HENAN UNIV OF SCI & TECH
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