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Food-grade recombined lactococcal lactis capable of producing conjugated linoleic acid as well as construction method and application thereof

A Lactococcus lactis, food-grade technology, applied in the field of food-grade recombinant Lactococcus lactis producing conjugated linoleic acid and its construction and application, can solve the problems of recombinant food-grade microorganisms that do not have high conversion efficiency of t10c12-CLA, etc.

Inactive Publication Date: 2013-06-12
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no reports of recombinant food-grade microorganisms with high conversion efficiency of t10c12-CLA

Method used

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  • Food-grade recombined lactococcal lactis capable of producing conjugated linoleic acid as well as construction method and application thereof
  • Food-grade recombined lactococcal lactis capable of producing conjugated linoleic acid as well as construction method and application thereof
  • Food-grade recombined lactococcal lactis capable of producing conjugated linoleic acid as well as construction method and application thereof

Examples

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Embodiment 1

[0042] Example 1, Construction of recombinant Lactococcus lactis with high conversion efficiency of t10c12-CLA

[0043]In this example, Lactococcus lactis (Lactococcus lactis) NZ3900 is used as the host bacterium, and pNZ8149-PAI is used as the recombinant food-grade expression vector as an example to illustrate the method of constructing recombinant Lactococcus lactis with high conversion efficiency of t10c12-CLA. In this method, after pNZ8149-PAI is transferred into Lactococcus lactis NZ3900 bacteria, the recombinant strains are screened by lactose medium, and the recombinant cells that can grow may carry the exogenous pai gene, which expresses the PAI isomerase Will convert linoleic acid to t10c12-CLA. This expression system avoids the use of antibiotics and does not produce endotoxin, and is an efficient food-grade vector expression system. Here, through the above experimental principles and methods, the pai gene was introduced into food-grade Lactococcus lactis through a...

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Abstract

The invention discloses food-grade recombined lactococcal lactis capable of producing conjugated linoleic acid as well as a construction method and an application thereof. The method for constructing the recombined lactococcal lactis comprises the step of introducing an encoding gene of a protein PAI (Plasminogen Activator Inhibitor) into lactococcal lactis serving as host bacteria to obtain the recombined lactococcal lactis, the conversion rate of the trans-10, cis-12 conjugated linoleic acid of which is higher than that of the host bacteria, wherein the protain PAI is a protein (a) composed of amino acid sequences shown in SEQ IN No. 2 or a protein (b) which is formed by replacing the amino acid sequences shown in SEQ IN No. 2 with one or several amino acid residues and / or deleting and / or adding the amino acid sequences shown in SEQ IN No. 2 and has the activity of linoleate isomerase, or is derived from the protein (1) and has the activity of linoleate isomerase. The food-grade recombined lactococcal lactis disclosed by the invention can produce t10c12-CLA, and the content of the t10c12-CLA accounts for 15.19-23.91% of the total quantity of thallus fatty acids.

Description

technical field [0001] The invention relates to food-grade recombinant Lactococcus lactis producing conjugated linoleic acid and its construction method and application. Background technique [0002] Conjugated linoleic acid (CLA) belongs to octadecadienoic acid and is an isomer of linoleic acid (LA), including more than 20 components. Biology of cis-9, trans-11 conjugated linoleic acid and trans-10, cis-12 conjugated linoleic acid (trans-10, cis-12; t10c12-CLA) isomers activity is relatively clear. For t10c12-CLA, it can promote the body's metabolism, inhibit fat synthesis, and reduce the probability of obesity. In 2008, the US FDA issued a food license, agreeing to use CLA as a food additive. At present, CLA has been produced and sold as a health product by many food and pharmaceutical companies. [0003] In nature, some rumen microorganisms in cattle and sheep have the ability to synthesize CLA, and these CLA can enter the milk. The total amount of CLA in one liter of...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/74C12P7/64C12R1/01
Inventor 苟克勉李世丽
Owner CHINA AGRI UNIV
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