Evaluation of fermentation potential of wild and UV-mutated yeasts screened from traditional murcha

J. Subba, B. Rai, D. Limbu, S. R. Rai
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Abstract

Murcha (an amylolytic starter) from different parts of Eastern Nepal were screened for fermentative yeasts. The most potential one was UV-mutated (8W lamp at λ = 254 nm and an intensity of 44.21 Wm-2 for 5-50 s) to study the effect of mutation on growth and fermentation properties. Respiratory-deficient mutants (RDMs) that resulted from the mutation were identified by triphenyl tetrazolium chloride (TTC) overlay technique and replica-plated for further isolation. Cell growth, substrate utilization, and ethanol yield of the mutants were compared with normal cells by carrying out fermentation in high-test cane molasses broth of 30 °Bx. An exhaustive screening of the samples resulted in only two murcha viz., from Laxmimarga (LM) and Udayapur (UD), having the desirable fermentation properties. UV-mutation study of UD and LM yeasts (both identified as strains of Saccharomyces cerevisiae) showed 8-12% survival and ~ 22% RDMs yield of the survived cells. Out of the 8 randomly selected RDMs, only UDm4 (colony No. 4 from UD) showed fermentation properties worth further investigation. Comparison of UD, LM and UDm4 by fermenting molasses (high test) broth of 30°Bx showed the least growth of UDm4 but the highest alcohol yield (9% and 16% more compared to UD and LM, respectively). The present finding indicates that it is possible to improve fermentation properties of feral yeasts from murcha by relatively simple UV-mutation approach. Finding the right mutant (the selective screening part), however, may involve considerable time and effort.
从传统murcha中筛选的野生和紫外突变酵母发酵潜力的评价
对尼泊尔东部不同地区的Murcha(一种解淀粉发酵剂)进行了发酵酵母筛选。利用8W紫外灯(λ = 254 nm,光照强度为44.21 Wm-2,光照时间为5 ~ 50 s)诱变菌株,研究其对菌株生长和发酵特性的影响。通过三苯基四氮氯化铵(TTC)覆盖技术鉴定了由该突变引起的呼吸缺陷突变体(rdm),并进行了复制镀。通过在30°Bx的高测试甘蔗糖蜜肉汤中进行发酵,将突变体的细胞生长、底物利用率和乙醇产量与正常细胞进行比较。对样品进行了详尽的筛选,结果只有两种murcha viz,来自Laxmimarga (LM)和Udayapur (UD),具有理想的发酵特性。对UD和LM酵母(均为酿酒酵母菌)进行紫外突变研究,发现存活细胞的存活率为8-12%,rdm产量为~ 22%。在8个随机选择的rdm中,只有UDm4(来自UD的4号菌落)显示出值得进一步研究的发酵特性。用30°Bx的糖蜜(高试)发酵液对UD、LM和UDm4进行比较,结果表明,UDm4的生长最少,但酒精产量最高(分别比UD和LM高9%和16%)。本研究结果表明,通过相对简单的紫外诱变方法可以改善野生murcha酵母的发酵特性。然而,找到合适的突变体(选择性筛选部分)可能需要花费大量的时间和精力。
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