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啤酒酵母和产朊假丝酵母属间原生质体融合子的筛选 总被引:2,自引:0,他引:2
利用不可逆生化抑制剂碘乙酸抑制一亲株细胞的生理活性和利用两亲株细胞间生理性状的差异性,进行啤酒酵母(Saccharomyces cerevisiae)和产朊假丝酵母(Candida utilis)原生质体融合子的筛选,属间原生质体融合率为3.47×10~(-6)。经菌落形态比较,碳化合物的同化和发酵,DNA含量测定,酯酶型分析,细胞核染色,产孢试验和自然的核分离实验证明,融合子分三种类型:87.21%产朊假丝醇母型;9.02%啤酒酵母型;3.77%真正核融合子型。 相似文献
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优良啤酒酵母原生质体融合株GR5的构建及其发酵特性 总被引:5,自引:0,他引:5
以发酵度较高的非絮凝性的啤酒酵母菌株X6和发酵度较低、絮凝性较强的啤酒酵母菌株N1为亲本进行原生质体融合。用亚硝酸诱变原养型的菌株X6,经筛选得到一株需酪素水解物的营养缺陷型菌株X6~20。采用正交试验法分别优化菌株X6~20和N1的原生质体形成和再生的条件。用X6~20菌株的原生质体作为受体和热灭活的N1菌株原生质体作为供体进行融合。融合株经三角瓶发酵筛选,得到一株较优良的融合株GR5。该融合株的絮凝性较强(本斯值为2.7),以12°Bx麦芽汁为培养基,用500 L的发酵罐在12℃下发酵,发酵至第8 d菌株GR5的发酵度为69.2%,发酵液中的双乙酰含量为0.0498 mg/L、乙醛含量为6.34 mg/L,总高级醇含量为74.4mg/L。融合株GR5具有双亲的优点,发酵的啤酒风味较好,是一株具有工业应用前景的啤酒酿造酵母菌株。 相似文献
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酿酒酵母与糖化酵母的种间原生质体融合及其融合子的鉴定 总被引:10,自引:0,他引:10
本文报道了酒精生产菌株K氏酿酒酵母Sacchormyces cerevisiae var.ellipsoideus的HUK-1(his-,二倍体,但在我们所试的5种产孢培养基上均不产孢)与糖化酵母Sac—charomyces diastaticus 7c(arg-,a)的种间原生质体融合,其营养标记互补的融合频率约是2.07×10-6—3.40×10-5。这些融合子曾在选择培养基MMs或Mmo上连续传代10次,以促进两亲核的融合。融合子的酒精发酵特性,细胞形态、体积大小、DNA含量、繁殖速率、发酵强度以及产孢能力等方面的观察和测定结果表明,均不同于双亲菌株。用显微操作器解剖了个别原养型融台子HU—KDF—185的4孢子子囊,在获得的93个单孢株中,其淀粉发酵特性和遗传标记均有双亲类型的分离或重组现象。上述实验结果充分证明了不同倍性的酵母之间可以通过原生质体融合获得种间杂种。 相似文献
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拉格啤酒酵母是我国啤酒酿造的主要菌种。细胞絮凝是啤酒酵母重要的生产性状,在不影响发酵性能的情况下适度提高酵母的絮凝能力,有助于发酵结束时细胞和产物的分离,有利于工业化啤酒生产,具有较高的经济价值。前期在对一株工业用拉格啤酒酵母G03及其絮凝突变株的研究中,挖掘到一个可能影响啤酒酵母絮凝性的候选基因RIM21。为了验证该基因的作用,文中在G03中对RIM21进行了敲除,发现RIM21敲除后,酵母在11 ℃发酵条件下的絮凝性能增强,基因FLO5、Lg-FLO1及细胞壁完整性途径中的部分基因表达上调。同时,CO2失重、酒精度、发酵度等发酵指标未有明显变化。另外,发现RIM21的缺失增强了啤酒酵母对细胞壁抑制剂的耐性。研究结果为阐释低温发酵条件下啤酒酵母的絮凝调控机理及菌株絮凝性的改善提供了基础。 相似文献
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用灭活的近裸香菇(Lentinus subnudus Berk.)双核菌株原生质体与香菇[L. edodes(Berk) Sing.] 双核菌株原生质体融合,在35℃的条件下选得融合子。融合频率为0—4.3x10-5。融合子与双亲有明显的拮抗性。融合子的菌丝形态、氨基酸含量,子实体的形态,以及酸性磷酸酶同功酶的测定都与双亲不同。 相似文献
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酵母菌属间原生质体融合构建高温酵母菌株 总被引:22,自引:0,他引:22
酿酒酵母(Saccharomyces cerevisiae) A001和克鲁维酵母(Kluyveromyces sp.) Y034属间原生质体融合构建高温酵母菌株。对制备高再生活性原生质体及融合子细胞形态、生理化特征、同工酶性质、遗传稳定性和高温发酵等方面进行了研究。结果表明,融合子AY023和AY680遗传性能稳定,表达了双亲优良性状,获得了在45℃培养条件下产酒率7.4%的属间融合菌株,是目前已见文献报道的产酒率最高的高温(45℃)酵母菌株。 相似文献
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克鲁维酵母种间原生质体融合的研究 总被引:5,自引:0,他引:5
乳酸克鲁维酵母(Kluyueromyces lactis Y12—1)和脆壁克鲁维酵母(K.fragilis8554)是乳糖酶生产菌株。应用原生质体融合技术进行了两菌株种问融合的研究。通过试验.原生质体形成及再生的最佳条件为:对数期的细胞,2%的蜗牛酶.30℃酶解30分钟.原生质体形成率90%以上,再生率20%左右。原生质体融合由聚乙二醇(PEG)诱导。K.lactisY12-l不能旋酵菊糖;K.fragilis 8554不能同化D-松三糖和麦芽糖;利用二菌株自身的营养缺陷性质获得融合子。融合子既能发酵菊糖又能同化D-松三糖和麦芽糖;融合子的DNA含量约为二亲株之和;融合子的菌落形态与亲株相比有一定差别.在以乳糖为碳源的培养基中,融合子的乳糖酶产量提高14一l6%;连续15次传代,融合子稳定。 相似文献
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K L Tr?ff R R Otero Cordero W H van Zyl B Hahn-H?gerdal 《Applied and environmental microbiology》2001,67(12):5668-5674
Saccharomyces cerevisiae ferments hexoses efficiently but is unable to ferment xylose. When the bacterial enzyme xylose isomerase (XI) from Thermus thermophilus was produced in S. cerevisiae, xylose utilization and ethanol formation were demonstrated. In addition, xylitol and acetate were formed. An unspecific aldose reductase (AR) capable of reducing xylose to xylitol has been identified in S. cerevisiae. The GRE3 gene, encoding the AR enzyme, was deleted in S. cerevisiae CEN.PK2-1C, yielding YUSM1009a. XI from T. thermophilus was produced, and endogenous xylulokinase from S. cerevisiae was overproduced in S. cerevisiae CEN.PK2-1C and YUSM1009a. In recombinant strains from which the GRE3 gene was deleted, xylitol formation decreased twofold. Deletion of the GRE3 gene combined with expression of the xylA gene from T. thermophilus on a replicative plasmid generated recombinant xylose utilizing S. cerevisiae strain TMB3102, which produced ethanol from xylose with a yield of 0.28 mmol of C from ethanol/mmol of C from xylose. None of the recombinant strains grew on xylose. 相似文献
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酿酒酵母工业菌株中XI木糖代谢途径的建立 总被引:9,自引:0,他引:9
根据代谢工程原理,采取多拷贝整合策略,利用整合载体pYMIKP,将来自嗜热细菌Thermusthermophilus的木糖异构酶(XI)基因xylA和酿酒酵母(Saccharomycescerevisiae)自身的木酮糖激酶(XK)基因XKS1,插入酿酒酵母工业菌株NAN-27的染色体中,得到工程菌株NAN-114。酶活测定结果显示,NAN-114中XI和XK的活性均高于出发菌株NAN-27,表明外源蛋白在酿酒酵母工业菌株中得到活性表达。对木糖、葡萄糖共发酵摇瓶实验结果表明,工程菌NAN-114消耗木糖4.6g/L,产生乙醇6.9g/L,较出发菌株分别提高了43.8%和9.5%。首次在酿酒酵母工业菌株中建立了XI路径的木糖代谢途径。 相似文献
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Molecular genetic identification of Saccharomyces sensu stricto strains from African sorghum beer 总被引:2,自引:0,他引:2
Genetic relationships of 24 phenotypically different strains isolated from sorghum beer in West Africa and the type cultures of the Saccharomyces sensu stricto species were investigated by universally primed polymerase chain reaction (PCR) analysis, microsatellite fingerprinting and PCR-restriction fragment length polymorphism (RFLP) of the ribosomal internal transcribed spacers. The results demonstrate that internal transcribed spacer (ITS) PCR-RFLP analysis with the endonucleases HaeIII, HpaII, ScrFI and TaqI is useful for discriminating S. cerevisiae, S. kudriavzevii, S. mikatae from one another and from the S. bayanus/S. pastorianus and S. cariocanus/S. paradoxus pairs. The sorghum beer strains exhibited the same restriction patterns as the type culture of S. cerevisiae CBS 1171. PCR profiles generated with the microsatellite primer (GTG)(5) and the universal primer N21 were almost identical for all isolates and strain CBS 1171. Despite phenotypic peculiarities, the strains involved in sorghum beer production in Ghana and Burkina Faso belong to S. cerevisiae. However, based on sequencing of the rDNA ITS1 region and Southern hybridisation analysis, these strains represent a divergent population of S. cerevisiae. 相似文献
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二氧化硫在啤酒中具有抗氧化的重要功能,而在其形成过程中APS激酶(MET14编码)起着非常重要的作用。以二氧化硫产量较高的青岛啤酒酵母(Saccharomyces cerevisiae)YSF-5的总DNA为模板,用PCR方法克隆得到MET14基因。为使目的基因在酿酒酵母中表达,以大肠杆菌-酿酒酵母穿梭质粒YEp352为载体,以PGK1强启动子为调控元件,构建了重组表达质粒pPM,并转化酿酒酵母YS58。转化子在YNB添加亮氨酸、组氨酸和色氨酸的选择性培养基上筛选鉴定,盐酸副玫瑰苯胺法测得转化子的SO2产量是受体菌的2倍左右。在重组表达质粒pPM的基础上添加铜抗性标记基因构建了重组表达质粒pCPM,并转化青岛啤酒工业酵母菌株YSF-38,转化子在YEPD 4mmol/L CuSO4的选择性培养基上筛选鉴定,实验室条件下培养后,测得转化子YSF-38(pCPM)的SO2产量是受体菌的3.2倍。用该转化子在青岛啤酒厂进行小型发酵实验,结果表明在发酵结束时,YSF-38(pCPM)转化子的SO2产量是受体菌的1.4倍。因此,MET14基因的有效表达可以提高啤酒工业酵母的SO2产量。 相似文献
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Mapping of Dbr1 and Ypk1 Suggests a Major Revision of the Genetic Map of the Left Arm of Saccharomyces Cerevisiae Chromosome Xi
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G. Simchen K. B. Chapman E. Caputo K. Nam L. Riles D. E. Levin J. D. Boeke 《Genetics》1994,138(2):283-287
The Saccharomyces cerevisiae dbr1 mutation has been mapped on the left arm of chromosome XI. XIL is a chromosome arm that was until now rather sparsely populated with accurately mapped markers. On the basis of physical data, the overall order of markers is inverted relative to the existing genetic map of XI. We present tetrad analyses using a variety of markers on XI that indicate that the existing genetic map of XIL should be inverted, at least for the strains in which our mapping was carried out, and probably for other S. cerevisiae strains. 相似文献
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近年来,随着发展低碳经济的迫切需要,可再生资源利用研究方兴未艾,其中,构建充分利用木质纤维素水解产物木糖生产乙醇的重组菌成为研究热点.木糖异构酶由于不需要辅酶,成为构建利用木糖重组酵母的首选途径.文中对近年来木糖异构酶的研究进展进行了综述.首先介绍了木糖异构酶的基本性质、序列、结构和功能特性,然后对其耐热机理进行了总结归纳;重点阐述了基于序列及结构进行的酶分子改造研究,包括底物特异性改造、热稳定性改造等;同时,结合作者的研究经历,对如何提高嗜热木糖异构酶在常温下的活性进行了探讨.最后,对木糖异构酶的研究进展进行了总结和展望,对基于结构改善木糖异构酶催化活性及构建新型高效利用木糖生产乙醇的重组菌具有重要的指导意义. 相似文献
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Electrophoretic karyotypes of yeast Saccharomyces cerevisiae integrant strains containing the pYF91 plasmid integrated into the chromosomes I, III, VI, IX, XI were studied. A possibility was demonstrated of visual identification of the chimaeric chromosomes via the molecular weight increase by 13200 bp (the plasmid size) determined by pulsed field gel electrophoresis. Several gamma-rays induced rearrangements of the yeast chimaeric chromosome I causing instability in hybrids were also studied. The deletions induced in the I chromosome were analysed and their size estimated. The technique of pulsed field gel electrophoresis is recommended for determination of insertions and deletions in the chromosomes of yeast Saccharomyces cerevisiae. 相似文献
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Four brewer's yeast strains carrying the alpha-ald gene of Klebsiella terrigena (ex. Aerobacter aerogenes) or of Enterobacter aerogenes on autonomously replicating plasmids were constructed. The alpha-ald genes were linked either to the ADC1 promoter or to the PGK1 promoter of yeast Saccharomyces cerevisiae. In pilot scale brewing (50 l) with three of these recombinant yeasts the formation of diacetyl in beer was so low during fermentation that lagering was not required. All other brewing properties of the strains were unaffected and the quality of finished beers was as good as that of finished beer prepared with the control strain. The total process time of beer production could therefore be reduced to 2 weeks, in contrast to about 5 weeks required in the conventional process. 相似文献
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Survival of Pectinatus cerevisiiphilus DSM 20466 in pure culture at variable temperatures under different oxygen concentrations was measured. Survival of P. cerevisiiphilus in co-culture with Saccharomyces cerevisiae under both saturated oxygen and brewing conditions was also studied. The survival of strictly anaerobic bacteria to oxygen seems to follow the classical laws of heat resistance. The D(oxy) values of P. cerevisiiphilus , calculated as a function of oxygen level, shows that the oxygen level is important for the survival duration of the bacteria. The temperature greatly influences the oxygen resistance of P. cerevisiiphilus, which increases when the temperature decreases. P. cerevisiiphilus resists better in co-culture than in pure culture under saturated oxygen conditions. Therefore, the oxygenation of the wort does not totally eliminate the risk of beer contamination by this bacterium. Under brewing conditions in co-culture at 8 degrees C, P. cerevisiiphilus grows slowly to reach a final cell concentration up to 10(6) cells/mL in beer, which is undrinkable. Pectinatus is a strictly anaerobic bacterium; however, it is resistant under certain oxygen conditions of incubation. This resistance is considerably higher in the presence of Saccharomyces cerevisiae . 相似文献