酮基还原酶在Pichia pastoris中表达策略

Study on strategy of keto reductase expression in Pichia pastoris

［目的］报道酮基还原酶在 Ｐｉｃｈｉａ ｐａｓｔｏｒｉｓ中的表达，探讨提高酮基还原酶表达量的策略．［方 法］用不同浓度的 Ｇ４１８甲醇、不同的 ｐＨ值、不同的诱导时间来考察对酮基还原酶表达水平的影响．［结 果］当 Ｇ４１８的浓度提高到 ３ ｇ／Ｌ，控制培养基的 ｐＨ在 ６～７之间，以甲醇与甘油之比为 １：１诱导 ４８ｈ时 酮基还原酶的表达水平最高，可达３０％

OBJECTIVE The expression of keto reductase in Pichia pastoris was reported, and the strategy for increasing the expression level of keto reductase was studied. METHODS Using different concentration of G418Methyl alcohol, different pH and different induced time, the expression level of keto reductase was reviewed. RESULTS The expression level of keto reductase was up to 30 % of total protein induced 48 hours by methanol glycerol(1: 1 ) under 3 g/L G418, medium pH 6-7.