A transformation of Yarrowia lipolytica and its construction method and application

Abstract

The invention discloses transformed yarrowia lipolytica, and a building method and application thereof. The transformed yarrowia lipolytica comprises a nucleotide sequence shown as SEQ ID NO:1, and is used for realizing the co-production of single-cell protein and lipase for feed; under the culture conditions that a YPD liquid culture medium is used, the liquid containing quantity is 10 percent, the rotating speed is 250rpm, the culture temperature is 28 DEG C, and the culture time is 84h, the protein content of the transformed yarrowia lipolytica is 60 to 70 percent, and the dry weight is 32g / L to 39g / L; the yield of the lipase for feed is 3200U / ml to 5900U / ml. The transformed yarrowia lipolytica is built by a gene engineering method; the co-production of the single-cell protein and the lipase for feed is realized; meanwhile, the content of two target products is increased.

Description

technical field [0001] The invention belongs to the field of biotechnology and feed industry, and more specifically relates to a transformation of Yarrowia lipolytica and its construction method and application. Background technique [0002] Lipase (lipase) is a typical carboxylic acid hydrolase, which is widely used in food, medicine, energy, environment and other fields because it can catalyze various reaction types such as hydrolysis, esterification, and transesterification. Compared with other feed enzymes, the application development is still in its infancy. The vast majority of microbial lipases are alkaline lipases, which have low tolerance to the acidic pH and digestive enzyme environment of the gastrointestinal tract of animals, and are difficult to exert catalytic activity in the gastrointestinal environment of animals. The feed lipase represented by Yarrowia lipolytica lipase has many characteristics required for feed enzymes, such as tolerance to gastrointestina...

AI Technical Summary

Problems solved by technology

[0003] Prior art can utilize biological carrier etc. to carry out the unit yield of lipase, as Yan Jinyong etc. utilize Pichia pastoris expression to produce lipase, as Wu Lan etc. utilize Yarrowia lipolytica to produce extracellular lipase; Yet its output is not high, lipase The yield is less than 1000U / ml, the dry weight of the bacteria is usually less than 20g / L, and the protein content is less than 50%

[0004] The existing technology usually adopts the optimization of physical and chemical processes or fermentation processes to increase the yield of the target product, but it can only achieve the improvement of a single target product, and the improvement range is limited