不同物理诱变方式对休哈塔假丝酵母产乙醇的影响
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摘要
采用不同物理方式(紫外线、He-Ne激光和微波)对休哈塔假丝酵母(Candida albicans)进行诱变,结合致死率、正突变率、乙醇产量等试验,分析不同诱变因子对该菌株发酵木糖的影响。结果表明,三种物理诱变对菌株诱变能力有差别,具有代表性的紫外线突变株(ZW-6)、He-Ne激光突变株(HN-3)及微波突变株(WB-3)的乙醇产量分别为17.44 g/L、18.49 g/L和18.11 g/L,较原始菌株分别提高13.91%、20.77%和18.29%;乙醇得率分别为0.35 g/g、0.38 g/g和0.37 g/g,较原始菌株分别提高13.80%、23.17%和19.50%。与紫外线及微波各诱变菌株相比,He-Ne激光诱变菌株的正突变率高,激光照射10 min时,正突变率高达48.62%;He-Ne激光诱变菌株的木糖利用率、乙醇产量、乙醇得率均提高最多,提高的范围最大,分别较原始菌株提高1.69%～44.77%、13.12%～40.59%、29.73%～47.75%。因此,确定He-Ne激光为最佳物理诱变因子。
Candida albicans was mutated by different physical methods(ultraviolet light, He-Ne laser and microwave), and combined with lethality,positive mutation rate, ethanol yield and other test results, the effects of different mutagenic factors on xylose fermentation were analyzed. The results showed that the three kinds of physical mutagenesis had different mutagenic ability, and the ethanol yields of representative UV mutants(ZW-6),He-Ne laser mutants(HN-3) and microwave mutants(WB-3) were 17.44 g/L, 18.49 g/L, and 18.11 g/L, respectively, which were 13.91%, 20.77%, and 18.29% higher than the original strain. The ethanol yields of the three strains were 0.35 g/g, 0.38 g/g, and 0.37 g/g, respectively, which were 13.80%,23.17%, and 19.50% higher than the original strain, respectively. Compared with the mutagenized strains of ultraviolet and microwave, the positive mutation rate of He-Ne laser mutagenized strain was high, and the positive mutation rate was as high as 48.62% after laser irradiation for 10 min; the xylose utilization rate, ethanol production and yield of He-Ne laser mutagenized strain increased most, which was 1.69%-44.77%, 13.12%-40.59%,29.73%-47.75% higher than the original strain, respectively. Therefore, it was determined that the He-Ne laser was the optimal physical mutagenesis factor.
Candida albicans was mutated by different physical methods(ultraviolet light, He-Ne laser and microwave), and combined with lethality,positive mutation rate, ethanol yield and other test results, the effects of different mutagenic factors on xylose fermentation were analyzed. The results showed that the three kinds of physical mutagenesis had different mutagenic ability, and the ethanol yields of representative UV mutants(ZW-6),He-Ne laser mutants(HN-3) and microwave mutants(WB-3) were 17.44 g/L, 18.49 g/L, and 18.11 g/L, respectively, which were 13.91%, 20.77%, and 18.29% higher than the original strain. The ethanol yields of the three strains were 0.35 g/g, 0.38 g/g, and 0.37 g/g, respectively, which were 13.80%,23.17%, and 19.50% higher than the original strain, respectively. Compared with the mutagenized strains of ultraviolet and microwave, the positive mutation rate of He-Ne laser mutagenized strain was high, and the positive mutation rate was as high as 48.62% after laser irradiation for 10 min; the xylose utilization rate, ethanol production and yield of He-Ne laser mutagenized strain increased most, which was 1.69%-44.77%, 13.12%-40.59%,29.73%-47.75% higher than the original strain, respectively. Therefore, it was determined that the He-Ne laser was the optimal physical mutagenesis factor.
引文
[1]黄达明,王美娟,管国强,等.运动发酵单胞菌和休哈塔假丝酵母混合发酵秸秆产乙醇[J].食品与发酵工业,2017,43(1):31-36.
[2]江枫,房伶晏,王浩,等.固定化混合菌利用戊糖和己糖共发酵制备乙醇[J].林业工程学报,2017,2(4):90-95.
[3]李晴,宋向阳,范晓阳,等.混菌戊糖己糖发酵产乙醇的研究[J].林产化学与工业,2015,35(2):111-116.
[4]蒋发现,黄萍,徐勇,等.稀酸预处理玉米秸秆共发酵产乙醇抑制物的来源探究[J].林产化学与工业,2015,35(4):85-91.
[5]余学军,张帆.在第十一届中国林业青年学术年会第8分会场上的讲话[C]//中国林业青年学术年会第8分会场论文集选.北京:中国林学会,2014:263-269.
[6]余恒,刘宝田,欧阳嘉.亚硫酸盐甘蔗渣浆酶解液发酵制备乙醇[J].生物加工过程,2013,11(5):7-10.
[7]刘红梅.多菌种混合共发酵甘蔗渣纤维素和半纤维素制备燃料乙醇的研究[D].广州:暨南大学,2015.
[8]GUO X,LIU Z,SONG Q,et al.Dynamics and mechanism of UV-damaged DNA Repair in indole-thymine dimer adduct:molecular origin of low repair quantum efficiency[J].J Phys Chem B,2015,119(8):3446-3355.
[9]吴雪昌,汪志芸,周婕,等.提高产抗生素链霉菌紫外诱变正变率的研究[J].遗传,2004,26(4):499-504.
[10]陈恒雷,徐辉,吕长武,等.激光诱变育种的研究概况[J].激光生物学报,2006,15(4):436-440.
[11]汪峰.氦氖激光器谐振腔调试实验的改进及研究[J].山东工业技术,2016(4):228.
[12]段良和,雷玉明,陈有为.氦氖激光对酒精酵母菌刺激效应的研究[J].昆明理工大学学报,1997,22(4):116-119.
[13]杜康,李广悦,丁德馨,等.产脲酶芽胞杆菌的微波诱变育种[J].微生物学杂志,2016,36(3):20-23.
[14]吴殿恺.微波诱变的实验研究与理论探讨[J].哈尔滨工业大学学报,1981(3):91-102.
[15]贾红华,周华,韦萍.微波诱变育种研究及应用进展[J].工业微生物,2003,33(2):46-50.
[16]葛菁萍,孙红兵,王轶男,等.氖激光和亚硝基胍复合诱变选育高产乙醇的休哈塔假丝酵母[J].激光生物学报,2011,20(3):292-298.
[17]MOHAGHEGHIA,EVANSK,CHOU Y C,et al.Cofermentation of glucose,xylose,and arabinose by genomic DNA-integrated xylose/arabinose fermenting strain of Zymomonas mobilis AX01[J].Appl Biochem Biotechnol,2002,98-100(1-9):885-898.
[18]宋莲芳.HPLC法测定木糖母液中的糖组分[J].安徽化工,2016,42(1):88-90.
[19]刘文信.HPLC在酒精发酵醪液检测中的应用[J].化学工程师,2013,27(2):28-30.
[20]MARIS A J A,WINKLER A A,KUYPER M,et al.Development of efficient xylose fermentation in Saccharomyces cerevisiae:Xylose isomerase as a key component[J].Biofuels,2007,108:179-204.
[21]杜仁鹏,黄守峰,裴芳艺,等.休哈塔假丝酵母木糖还原酶基因(xyl1)的克隆与序列分析[J].中国农学通报,2015,31(11):124-129.
[2]江枫,房伶晏,王浩,等.固定化混合菌利用戊糖和己糖共发酵制备乙醇[J].林业工程学报,2017,2(4):90-95.
[3]李晴,宋向阳,范晓阳,等.混菌戊糖己糖发酵产乙醇的研究[J].林产化学与工业,2015,35(2):111-116.
[4]蒋发现,黄萍,徐勇,等.稀酸预处理玉米秸秆共发酵产乙醇抑制物的来源探究[J].林产化学与工业,2015,35(4):85-91.
[5]余学军,张帆.在第十一届中国林业青年学术年会第8分会场上的讲话[C]//中国林业青年学术年会第8分会场论文集选.北京:中国林学会,2014:263-269.
[6]余恒,刘宝田,欧阳嘉.亚硫酸盐甘蔗渣浆酶解液发酵制备乙醇[J].生物加工过程,2013,11(5):7-10.
[7]刘红梅.多菌种混合共发酵甘蔗渣纤维素和半纤维素制备燃料乙醇的研究[D].广州:暨南大学,2015.
[8]GUO X,LIU Z,SONG Q,et al.Dynamics and mechanism of UV-damaged DNA Repair in indole-thymine dimer adduct:molecular origin of low repair quantum efficiency[J].J Phys Chem B,2015,119(8):3446-3355.
[9]吴雪昌,汪志芸,周婕,等.提高产抗生素链霉菌紫外诱变正变率的研究[J].遗传,2004,26(4):499-504.
[10]陈恒雷,徐辉,吕长武,等.激光诱变育种的研究概况[J].激光生物学报,2006,15(4):436-440.
[11]汪峰.氦氖激光器谐振腔调试实验的改进及研究[J].山东工业技术,2016(4):228.
[12]段良和,雷玉明,陈有为.氦氖激光对酒精酵母菌刺激效应的研究[J].昆明理工大学学报,1997,22(4):116-119.
[13]杜康,李广悦,丁德馨,等.产脲酶芽胞杆菌的微波诱变育种[J].微生物学杂志,2016,36(3):20-23.
[14]吴殿恺.微波诱变的实验研究与理论探讨[J].哈尔滨工业大学学报,1981(3):91-102.
[15]贾红华,周华,韦萍.微波诱变育种研究及应用进展[J].工业微生物,2003,33(2):46-50.
[16]葛菁萍,孙红兵,王轶男,等.氖激光和亚硝基胍复合诱变选育高产乙醇的休哈塔假丝酵母[J].激光生物学报,2011,20(3):292-298.
[17]MOHAGHEGHIA,EVANSK,CHOU Y C,et al.Cofermentation of glucose,xylose,and arabinose by genomic DNA-integrated xylose/arabinose fermenting strain of Zymomonas mobilis AX01[J].Appl Biochem Biotechnol,2002,98-100(1-9):885-898.
[18]宋莲芳.HPLC法测定木糖母液中的糖组分[J].安徽化工,2016,42(1):88-90.
[19]刘文信.HPLC在酒精发酵醪液检测中的应用[J].化学工程师,2013,27(2):28-30.
[20]MARIS A J A,WINKLER A A,KUYPER M,et al.Development of efficient xylose fermentation in Saccharomyces cerevisiae:Xylose isomerase as a key component[J].Biofuels,2007,108:179-204.
[21]杜仁鹏,黄守峰,裴芳艺,等.休哈塔假丝酵母木糖还原酶基因(xyl1)的克隆与序列分析[J].中国农学通报,2015,31(11):124-129.