CAJ | 학술논문

以啤酒酵母G-03为模板,扩增得到铜抗性基因(cup1)和β-葡聚糖合成酶基因(fks1).将fks1连接pMD-18T Vector得到重组质粒pTK,重组质粒pTK和cup1经BglⅡ、SalⅠ酶切后连接得到重组质粒pKC. BamHⅠ酶切重组质粒pKC 得到以fks1为整合位点包含cup1的基因片段fks1::cup1.用此片段转化啤酒酵母工业菌株G-03,通过硫酸铜抗性筛选得到一株啤酒酵母工程菌G-03/C.工程菌连续传代10次后依然能在筛选平板上生长,遗传稳定性良好.主酵结束G-03/C和G-03的辛酸和癸酸含量基本相同.25 ℃诱导自溶20 d,G-03/C的辛酸、癸酸分别下降57.3%、81.8%,自溶性能减弱.G-03/C的死亡率、双乙酰、浊度及TBA均较原菌有所下降.G-03/C与G-03酿制成品啤酒的常规指标没有较大差别,品评结果表明,G-03/C风味更优.
이비주효모G-03위모판,확증득도동항성기인(cup1)화β-포취당합성매기인(fks1).장fks1련접pMD-18T Vector득도중조질립pTK,중조질립pTK화cup1경BglⅡ、SalⅠ매절후련접득도중조질립pKC. BamHⅠ매절중조질립pKC 득도이fks1위정합위점포함cup1적기인편단fks1::cup1.용차편단전화비주효모공업균주G-03,통과류산동항성사선득도일주비주효모공정균G-03/C.공정균련속전대10차후의연능재사선평판상생장,유전은정성량호.주효결속G-03/C화G-03적신산화계산함량기본상동.25 ℃유도자용20 d,G-03/C적신산、계산분별하강57.3%、81.8%,자용성능감약.G-03/C적사망솔、쌍을선、탁도급TBA균교원균유소하강.G-03/C여G-03양제성품비주적상규지표몰유교대차별,품평결과표명,G-03/C풍미경우.
A copy of copper response gene (cup1) and a copy of β-glucan synthetase gene(fks1) were extended by PCR reaction from the industrial brewing yeast strain G-03. Recombinant plasmid pKC was constructed by linking cup1 to pTK, which was made up of fks1 and pMD-18T Vector, by restrict enzyme BglⅡ、SalⅠ. The fragment fks1::cup1 obtained through the restrictive enzyme reaction was integrated to the chromosomal DNA of G-03 strain, and recombinants were screened by copper resistance. To test the genetic stability, strain G-03/C was incubated into flasks and transferred continuously 10 times. It could grow on copper resistant medium. When the main fermentation was over, there was no differences in the concentration of C_(8:0) and C_(10:0) between G-03 and G-03/C. Under 25 degree forced autolytic conditions, the content of fatty acids including C_(8:0), C_(10:0), reduced by 57.3%, 81.8% respectively, compared with that of the host strain G-03. Therefore the autolytic ability weakened, while mortality, concentration of diacetyl, turbidity and TBA of G-03/C reduced to a certain extent. Sensory evaluation test of beer demonstrated that the beer flavor of G-03/C was better.