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Thermomyces lanuginosa lipase gene, engineering bacteria and its application

A technology of thermomyces and lipase gene, which is applied in the fields of genetic engineering, application, plant gene improvement, etc., can solve the problems of application limitation and high cost, and achieve the goal of improving stability, high-efficiency expression, and improving translation initiation efficiency Effect

Active Publication Date: 2018-06-26
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the high cost of sn-1,3-specific lipase limits its further application

Method used

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  • Thermomyces lanuginosa lipase gene, engineering bacteria and its application
  • Thermomyces lanuginosa lipase gene, engineering bacteria and its application
  • Thermomyces lanuginosa lipase gene, engineering bacteria and its application

Examples

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Effect test

Embodiment 1

[0035] (1) Improvement of the lipase coding gene of Thermomyces lanuginosa

[0036] The existing disclosed Thermomyces lanuginosa lipase coding gene is shown in SEQ ID NO.3, and the gene is derived from Thermomyces lanuginosa, which is different from yeast, which may be the cause of the The reason why the expression level of the gene is not high in Pichia pastoris.

[0037] In this example, on the basis of the amino acid sequence (SEQ ID NO.1) of Thermomyces lanuginosa lipase encoding (TLL), a codon preferred by Pichia pastoris was used to replace the low frequency in the RML coding gene in Pichia pastoris The codons used (as shown in Table 1), while removing factors that are not conducive to protein expression such as strong stem-loop structures and potential early termination signals, designed a gene sequence suitable for high-efficiency expression in Pichia pastoris, The specific sequence is shown in SEQ ID NO.2, so as to improve the expression level of Thermomyces lanugin...

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Abstract

The invention discloses a thermomyces lanuginosus lipase gene, engineering bacteria and an application of the engineering bacteria. The base sequence of the thermomyces lanuginosus lipase gene is shown in SEQ ID NO.2. On the basis that existing thermomyces lanuginosus lipase amino acid sequences and protein structures are not changed, factors adverse to protein expression, for example, rare codons, rare codon pairs, strong stem-loop structures, potential early termination signals and the like are avoided, and the thermomyces lanuginosus lipase gene sequence is optimized, so that the sequence can efficiently express thermomyces lanuginosus lipase in a pichia pastoris recombination expression system, and the expression vitality is up to 370.65 U / ml and is improved by 58.38% while compared with the recombination expression vitality of the original sequence of the thermomyces lanuginosus lipase gene being 234.03 U / ml.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a thermomyces lanuginosa lipase gene, engineering bacteria and applications thereof. Background technique [0002] Lipase (E.C.3.1.1.3), also known as acylglycerol hydrolase, mainly catalyzes the hydrolysis of triacylglycerides in nature, and can also catalyze esterification, transesterification, alcoholysis and enzymatic hydrolysis under non-aqueous conditions reaction. In addition, lipases have various substrate specificities, such as fatty acid-specific lipases, position-specific lipases, and stereospecific lipases. The catalytic diversity and substrate specificity of lipase make it widely used in many fields such as grain and oil production, food industry, daily chemical industry, oleochemical industry, agricultural chemical industry, paper industry, detergent industry and drug synthesis. application. [0003] sn-1,3, position-specific lipase, has great application ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/55C12N15/81C12N1/19C12N9/20C12R1/84
Inventor 冯凤琴蔡海莺赵敏洁李杨张辉
Owner ZHEJIANG UNIV
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