正十六烷发酵产生柠檬酸的研究

通过试验筛选出产柠檬酸较高的菌种解脂复膜孢酵母１４６０菌和适宜培养基,确定了适宜的菌种起始浓度和底物浓度,证实谷氨酸对产生柠檬酸的促进作用,对分批培养发酵过程中细胞生长和产物生成速率进行了数学表达。     

扣囊复膜酵母在红枣酒中的应用

采用纯培养技术对10个采集自大竹县东柳乡的米酒曲样品蕴含的酵母菌进行分离鉴定,结果表明,24株酵母菌中16株被鉴定为Saccharomycopsis fibuligera(扣囊复膜酵母),S.fibuligera为米酒曲中的优势酵母菌。选取S.fibuligera分离株制备红枣酒并对其品质进行评价,结果发现红枣酒酒精度在8.1%vol~11.9%vol,可溶性固形物含量在7.59%~17.21%。色度仪结果表明,红枣酒颜色整体偏红偏黄。电子鼻结果表明,乙醇为红枣酒挥发性化合物的主要成分。电子舌结果表明,红枣酒在酸味指标上差异最大,相对强度的极差值为4.48。主成分分析结果表明,S.fibuligera HBUAS61136酿造的红枣酒具有较好的风味品质和较高的酒精度。     

正十六烷发酵产生柠檬酸的研究

通过试验筛选出产柠檬酸较高的菌种解脂复膜孢酵母１４６０菌和适宜培养基，确定了适宜的菌种起始浓度和底物浓度，证实谷氨酸对产生柠檬酸的促进作用。     

Cloning, sequencing and characterization of the alpha-aminoadipate reductase gene (LYS2) from Saccharomycopsis fibuligera

The gene putatively encoding alpha-aminoadipate reductase (AAR) was isolated successfully by degenerate PCR and chromosome walking, based on cassette PCR methods, from the dimorphous yeast Saccharomycopsis fibuligera PD70 and was named SfLYS2. Sequence analysis revealed that it contained a putative open reading frame (ORF) of 4161 bp and encoded a polypeptide of 1386 amino acids. The deduced translation product shared an identity of 53% and 51% to the Lys2p homologues of Candida albicans and Saccharomyces cerevisiae, respectively. An atypical TATA box and a GCN4-box element were found in the 5'-upstream region. Genomic Southern hybridization suggested the presence of a single locus of SfLYS2 in the S. fibuligera genome. Expression of the ORF of SfLYS2 in a lys2(-) strain of S. cerevisiae could functionally complement the lysine mutant of the S. cerevisiae strain. S. fibuligera could use lysine as the sole nitrogen source but its growth was inhibited on the alpha-aminoadipate (AA) medium. Approximately 90% of the mutants of S. cerevisiae resistant to AA are lysine auxotrophs; in contrast all the mutants of S. fibuligera resistant to AA recovered in this work were not lysine auxotrophs.     

高产酸扣囊复膜酵母的筛选与培养基配方优化

该研究对10种不同的米酒曲和黄酒曲中的高产酸酵母菌进行了分离、筛选及鉴定,并以酵母菌的生物量为评价指标,采用单因素试验和响应面法,对菌株的液态发酵培养基进行优化。结果表明,经过分子生物学鉴定,共分离获得6株扣囊复膜酵母（Saccharomycopsis fibuligera）。通过菌株产酸能力和耐受性的比较,筛选到一株产酸率高、耐高温和乙醇能力强的扣囊复膜酵母菌株3-1,其总酸（以乳酸计）产量达5.4 g/L。最佳培养基配方为：糖蜜7.5 g/L,葡萄糖7.7 g/L,大豆蛋白胨1.7 g/L,酵母浸粉1.7 g/L。在此优化条件下,菌株3-1的生物量达2.45×108个/mL,总菌数比对照培养基提高了63.3%。     

甜酒酿中酵母菌多样性的研究

从福建酒曲、苏州酒曲和安琪酒曲中分离出6株酵母菌.通过对所分离的酵母菌的18S rDNA进行分析鉴定,同时利用形态、生理生化的分析,最终确定这6株菌属于二个类群,其分别属于酿酒酵母属,复膜酵母属.利用18S rDNA的序列,进行聚类分析,这6株酵母菌其中福建酒曲中有4株酵母菌,分别为F J-2、F J-5、F J-6、F J-7,其中FJ-2与Saccharomyces cerevisiae strain OC 11的相似性达到99％、F J-5与S.cerevisiae S288c相似性达99％、F J-6与S.cerevisiae strain OC11相似性达99％,F J-7与Saccharomycopsis fibuligera strain YW12相似度达到99％.苏州酒曲中有1株,为SZ-2与S.cerevisiae的相似性达99％.安琪酒曲中有1株,该菌株与酿酒酵母的相似度为100％.由此可见,在酒酿中发挥作用的酵母菌是酿酒酵母属和复膜酵母属中的菌株.     

产黄青霉、扣囊复膜孢酵母替代部分大曲提高食醋原料利用率

从分离自山西老陈醋大曲中高产糖化酶和纤维素酶的菌株中筛选优良菌株,制成麸曲与大曲共发酵,采用混料回归设计确定最佳原料配比并检测食醋质量。结果表明,以菌株AS3.4309（M0）为对照菌株,筛选出产黄青霉（M4）和扣囊复膜孢酵母（M8）,大曲与麸曲的最佳总添加量为主料的40%,其中大曲25.10%,M8麸曲6.81%,M4麸曲8.09%,安琪酵母0.06%,料水比1∶3（g∶mL）,此时酒精度为11.2%vol。制成麸曲与大曲共发酵后酒精度显著高于菌株M0（P＜0.05）,与大曲单独发酵相比,麸曲M4+M8+大曲共酵提高原料利用率提高了31.81%,减少大曲用量37.25%,且食醋各项质量指标均符合国家标准。     

Glycerol accumulation in the dimorphic yeast Saccharomycopsis fibuligera: cloning of two glycerol 3-phosphate dehydrogenase genes, one of which is markedly induced by osmotic stress

Glycerol plays an important role in the osmoadaptation responses of Saccharomyces cerevisiae. However, there is no detailed investigation about the role of glycerol in the osmoadaptation responses of Saccharomycopsis fibuligera. Here we show that both intra- and extracellular glycerol concentrations in Sm. fibuligera cells responded very quickly when they were subjected to osmotic stress. We then cloned two isogenes encoding putative NAD(+)-dependent glycerol 3-phosphate dehydrogenase (GPD) from Sm. fibuligera PD70 by degenerate PCR and subsequent chromosome walking methods. Those two genes, designated SfGPD1 and SfGPD2, respectively, exhibited 86.6% pairwise identity in their encoding regions, while there was no obvious homology in their non-coding regions. Either SfGPD1 or SfGPD2 could complement the salt tolerance characteristics of the gpd1gpd2 double mutant strain of S. cerevisiae, further demonstrating that both of those genes are functional homologues of S. cerevisiae GPD1 and GPD2. Northern blot analysis revealed that SfGPD1 was induced markedly by osmotic stress, while SfGPD2 was not. In consistency with the observation that there was no obvious glycerol content change when the cells were transferred to anoxic conditions, neither SfGPD1 nor SfGPD2 was induced when the cells were transferred to anoxic conditions, thus suggesting a functional splitting of glycerol 3-phosphate dehydrogenase between S. cerevisiae and Sm. fibuligera.     

烧酒曲中扣囊复膜酵母的分离及鉴定

为了筛选1株可用于纯菌种酿造糯米酒的酵母菌,将烧酒曲稀释涂布在淀粉培养基(YPS)平板上培养,得到1株产淀粉酶酵母菌(YW12)。通过形态学、生理生化特征和18S rDNA、ITS区序列分析鉴定,YW12为扣囊复膜酵母(Saccharomycopsis fibuligera)。YW12在YPS液体培养基中28℃培养4 d,用Yoo改良法测其粗酶液的淀粉酶活力为49.8 U/mL。YW12在糯米糖化液(含175 g/L葡萄糖)中28℃发酵3 d的酒精度为5.63%(v/v)。YW12既能同化淀粉又能发酵产生酒精,具有用于纯菌种酿造糯米酒的潜力。     

Effects of the proteins of indica rice and indica waxy rice on the formation of volatiles of sweet rice wine

The proteins in the raw materials affect the formation of flavour during fermentation. Among all the fermentation broth of protein fraction in the study, the rice prolamin and albumin effectively promoted floral, honey and fruity characteristics in fermentation with higher level of β-phenylethyl alcohol, 2,4-dimethyl-benzaldehyde and ethyl hexadecanoate than those in the fermentation of globulin and glutelin; albumin is with higher level of higher alcohols, but higher alcohols are not good for health because of their acute toxicity and neurotoxic effects. In a further fermentation experiment of addition of prolamin, extra prolamin significantly promoted the formation of aroma volatiles, especially medium and long fatty acid esters and β-phenylethyl alcohol and its acetate, and the content of β-phenethyl acetate in sweet rice wine with added prolamin was more than twice that of single sweet rice wine. This study demonstrates that the different rice protein components significantly affect the volatiles generated by microorganism metabolism to impact the flavour of sweet rice wine, and the flavour quality of sweet rice wine can be enhanced by increasing the prolamin content.