The Cloning Of Aldehyde Dehydrogenase4Gene Of Saccharomyces Cerevisiae As2.399and The Optimization Of Fermentation Medium In S. Cerevisiae

Aldehyde dehydrogenase is a kind of enzyme which can convert alcohols in organisms into the corresponding acids, widely existing in animals, plants, micro-organisms. In the human body, ALDH is a key metabolic enzymes, which can metabolize aldehyde which alcohol can be converted into by alcohol dehydrogenase, to acetic acid, and then the acetic acid will be changed into water and carbon dioxide, or use for the synthesis of other nutrients in vivo. In plants, aldehyde dehydrogenase is mainly induced in the stress environment, in order to help the plant itself against environmental stress, achieving the purpose of survival. In the microorganism, the function of aldehyde dehydrogenase is mainly to cope with environmental stress, seeking to live space.There is many kinds of aldehyde dehydrogenase in Yeast, up to now, six kinds of aldehyde dehydrogenase have been found, ALD1-ALD6. They have different distributions and expression ways in the yeast, the ALD4and ALD5distributed in mitochondrion metabolize aldehyde which is transported from cytoplasm to the mitochondrion.ALD1, ALD2, ALD3, and ALD6are mainly distributed in the yeast cytoplasm and complete the corresponding function.This experiment mainly was divided into two parts:1. The experiment have targeted gene ALD4from Saccharomyces cerevisiae AS2.399cloned into Pichia pasoris GS115by TA and electrotransformation, constructing engineering bacteria P.pasoris GS115(Ppic9K-ALD4).2. By orthogonal experiment, we gained a high density fermentation culture medium of Saccharomyces cerevisiae AS2.399.As the same time,the ethanol, sorbitol,and ionic liquids were used as inducers,in order to induce the expression of acetaldehyde dehydrogenase.