CAJ | 학술논문

目的：获得丙三醇高产多形汉逊酵母重组菌株。方法：利用低能氮离子（N+）介导转基因技术介导多形汉逊酵母基因组总DNA随机转化自身，构建丙三醇高产工程菌。通过高盐培养基初筛，纸色谱法和气相色谱法复筛目标菌株。进而运用正交试验对发酵培养基进行优化。结果：获得一株遗传稳定的能够高效生产丙三醇的多形汉逊酵母重组菌株，其产量高达70.29g/L。对重组菌株传代培养8代，利用气相色谱分析发现丙三醇产量基本稳定。结论：利用低能N+离子介导多形汉逊酵母基因组转化自身宿主技术，可以在丙三醇生物合成及相关调控基因未知的情况下构建丙三醇高产的多形汉逊酵母重组菌株。
목적：획득병삼순고산다형한손효모중조균주。방법：이용저능담리자（N+）개도전기인기술개도다형한손효모기인조총DNA수궤전화자신，구건병삼순고산공정균。통과고염배양기초사，지색보법화기상색보법복사목표균주。진이운용정교시험대발효배양기진행우화。결과：획득일주유전은정적능구고효생산병삼순적다형한손효모중조균주，기산량고체70.29g/L。대중조균주전대배양8대，이용기상색보분석발현병삼순산량기본은정。결론：이용저능N+리자개도다형한손효모기인조전화자신숙주기술，가이재병삼순생물합성급상관조공기인미지적정황하구건병삼순고산적다형한손효모중조균주。
Objective:To obtain recombinant Hansenula polymorpha strain with high yield of glycerol. Methods:We have developed an efficient method to construct recombinant Hansenula polymorpha strains with enhanced ability to produce glycerol by randomly incorporating total genomic DNA of Hansenula polymorpha into itself through low-energy nitrogen ions (N+)-mediated genetic transformation. To screen efficiently target recombinant strains, a tolerant to high salt culture-based high-throughout screening was utilized. Then paper chromatography and gas chromatography were used to further analyze the candidate strains based on the preliminary screening. Additional y, the orthogonal experiment was used to optimize the fermentation medium. Results:One strain of genetical y stable recombinant Hansenula polymorpha strain capable of yielding efficiently glycerol was constructed, and its yield is up to 70.29g/L. Meanwhile, the recombinant strain was subcultured for eight generations, and the output is stable based on the GC analysis. Conclusion:Hansenula polymorpha total genomic DNA can be transformed randomly into itself using low-energy nitrogen ions (N+)-mediated genetic transformation to obtain recombinant Hansenula polymorpha strains with high-yield of glycerol when the biosynthesis of glycerol is stil unknown .