Genome shuffling技术选育高耐性酿酒酵母

酿酒酵母对温度和乙醇的耐受性对于茅台酒糟再利用工艺是至关重要的.通过紫外诱变方法和筛选,获得了5个耐性有所提高的正突变株,以这些菌株作为出发菌,进行连续融合,复合筛选融合子,最终获得了能耐受46℃和16%vol乙醇浓度的酵母菌.耐高温和耐酒精能力分别提高了7%和33%.经过摇瓶发酵实验后证明.该菌株在30℃下发酵4 d酒精度达13.87%vol,总酯产量为0.637 g/L,42℃下发酵4 d酒精度达5.30%vol,总酯产量为0.313g/L.     

耐酸性运动发酵单胞菌的筛选和酒精发酵条件的优化

通过酸性平板筛选,在作者所在实验室保藏的运动发酵单胞菌中,得到一株能在pH4.5时进行酒精发酵的菌株,其酒精产量为66.7g/L,与原菌株在pH6.5时的相同.通过单因素实验和正交实验,对该菌在酸性条件下的酒精发酵条件进行了优化.结果表明,在发酵温度为30～35℃,pH4.5,初糖质量浓度150g/L,酵母膏质量浓度4g/L,蛋白胨质量浓度为2g/L时,该菌的酒精产量达到最大,为71.7g/L.     

发酵木糖酵母菌株的选育

本文介绍了发酵木糖产酒精酵母菌种的筛选方法并利用该方法筛选出一株性能优良的发酵木糖产酒精酵母菌株Z8。该菌株性能测试结果为 :发酵木糖产乙醇能力相当于理论产量的 60 .0 % ;发酵葡萄糖浓度为 8%的发酵液 ,乙醇转化率为0 .3 88g/g;最低乙醇抑制浓度 14% (v/v) ;在温度高达 42℃的条件下仍能正常生长。并初步鉴定该菌株为奥默毕赤氏酵母(Pichiaahmeri)。     

增强海藻糖胞内积累提高大肠杆菌耐受性与乙醇产率

为提高产乙醇大肠杆菌对发酵底物和乙醇的耐受性,并提高乙醇发酵性能,以大肠杆菌B0013-1031H为出发菌株,对其海藻糖代谢途径进行改造,获得了敲除海藻糖分解途径的突变株JC31和进一步加强海藻糖合成途径的突变株JC41。突变株JC31和JC41较出发菌株都具有更高的海藻糖合成与积累能力,其中JC41的胞内海藻糖含量可达出发菌株的12倍。与出发菌株相比,突变株JC31和JC41对葡萄糖和乙醇胁迫的耐受性显著提高。进一步引入乙醇合成途径,在葡萄糖质量浓度120 g/L的发酵条件下,菌株JC31-PA表现出最优的发酵性能,其最大乙醇产量为50. 6 g/L,较对照菌株提高了5. 42%;乙醇转化率为48. 72 g/100 g葡萄糖,较对照提高了12. 67%,达到理论转化率的95%。     

高菊粉酶活酿酒酵母的诱变选育

以安琪酿酒酵母为出发菌株,采用紫外与微波2种物理诱变方法进行复合诱变.经过初筛与复筛,选育出了高菊粉酶活突变菌株Y05,其酶活达到90.97U/mL,约为出发菌株的8倍.经多次传代证明该菌株遗传稳定性良好.当培养基中菊粉浓度200g/L时,经30℃72h发酵突变菌株Y05发酵液乙醇浓度为71.78g/L,乙醇得率为0.43g/g,分别比出发菌株提高了42.5%和13.2%.     

耐酸性运动发酵单胞菌酒精发酵条件的初步研究

在本实验室保藏的运动发酵单胞菌中 ,通过化学诱变和酸性平板分离 ,得到一株能在pH 4 5时进行酒精发酵的菌株 ,其酒精产量为 6 6 7g/L ,与原菌株在pH 6 5时的 6 6 7g/L相同。研究了种龄、接种量、发酵初始pH和发酵温度对细菌酒精发酵的影响。结果表明 ,在种龄为 12hr,接种量 10 % ,发酵温度 30℃～ 35℃ ,pH 4 5时 ,该菌的酒精产量达到最大为 70 1g/L。     

耐高温高产酒精酵母菌的筛选鉴定及特性研究

为了筛选耐高温产酒精酵母菌,以贵州省茅台镇酱香型酒厂的第四轮次酒醅为实验材料,采用平板划线分离法筛选出37株酵母菌,从37株酵母菌中通过热致死温度、TTC染色法获得1株耐高温且高产酒精的J18菌株,对其在最高发酵温度的耐乙醇、耐酸、耐糖及发酵力进行了研究。结果表明：J18菌株的最高发酵温度为44℃,耐乙醇浓度18%vol,最适生长pH值为5.5;最适生长葡萄糖浓度为35%,最适发酵条件下发酵力为4.881 g/100 mL。对该菌株进行形态学鉴定和26S rDNA测序,确定该菌株为酿酒酵母（Saccharomyces cerevisiae）。     

富硒酵母的选育及其发酵条件的优化试验

通过对一株耐硒酵母进行UV和60Co逐级诱变,获得一株产量高的突变株Y7,发酵液的硒总含量达19mg/L,是出发菌株的1.58倍,经多次传代试验,证明其稳定性良好。通过单因素、L16(37)正交试验对其发酵培养基和培养条件进行优化,试验表明富硒酵母发酵培养基的最佳配方是:蔗糖6%,牛肉膏1%,蛋白胨1%,K2HPO4 0.15%,亚硒酸钠35μg/L;最佳培养条件:初始pH为4.0,温度32℃,装液量60mL/250mL摇瓶,转速200r/min,10%(v/v)接种量,培养48h。富硒酵母的硒总含量达23mg/L以上,是出发菌株的1.92倍。     

紫外诱变选育耐酸酵母菌株及其特性研究

以实验室保存的耐酸酵母菌株YM1-1为出发菌株,在其对数生长期进行紫外诱变处理,经过三级筛选后,最终筛得一株在pH 1.8强酸环境中仍可存活的突变菌株YM1-1E。在其特性研究中发现,菌株YM1-1E最适生长温度和pH值分别为36 ℃和5.5,能耐受500 mg/L SO2、14%乙醇和65%的葡萄糖溶液。突变菌株YM1-1E在pH值为2.5的发酵培养基中发酵6 d后,发酵液酒精度为4.5%vol,糖醇转化率为46.39%,可溶性固形物含量为9.7%,较出发菌株YM1-1相比,产酒精能力提升了25%。     

细菌纤维素产生菌选育与突变株发酵特性的研究

为了获得细菌纤维素高产菌株,采用紫外线和硫酸二乙酯对实验室保存的木葡糖酸醋杆菌BC13进行复合诱变。通过诱变、筛选和连续传代实验,获得1株产量达15.6g/L的高产突变株Y19-11,该菌株遗传稳定性良好,比原出发菌株产量提高44.4%。同时,通过比较不同温度、不同初始pH下的菌体量和细菌纤维素产量,确定突变株Y19-11的最适温度和初始pH分别为30℃和5.5。     

橘园分离的非酿酒酵母的耐受性分析

为了更好地利用从橘园分离到的10株非酿酒酵母酿造果酒,对这10株非酿酒酵母的发酵力,糖、酒精和SO2的耐受性进行分析。结果显示,不同种的酵母菌发酵力不同,对酒精、糖和SO2耐受性有很大差异,其中酒精对酵母菌的抑制作用最大。2株酿酒酵母在这些耐受条件下表现良好,虽然有抑制作用,但依然能够正常生长。自分离的10株非酿酒酵母中,Candida tropicalis 和C. humilis 发酵力最高,Issatchenkia orientalis 和Torulaspora delbrueckii 能够耐受8%vol的酒精,C. tropicalis,C. humilis 和T. delbrueckii 能够耐受260 g/L的葡萄糖,180 mg/L的SO2。     

Leuconostoc fallax,an acid and ethanol tolerant lactic acid bacterium

Leuconostoc fallax, known to be present in sauerkraut, was reisolated from exudates of Gerbera jamesonii. The identity of the isolates with L fallax was demonstrated by sequence analysis of the first 600 bases of the 16S rRNA. L fallax utilised a small number of sugars and showed a remarkable resistance to lactic acid. The final pH in glucose broth was 3·9. Moreover it was able to grow in the presence of ethanol (9·0% v/v) and salt (5·5%), but it was unable to carry out a malo-lactic fermentation.     

酿酒活性干酵母生理特性的研究

建立一种酿酒活性干酵母生理活性的简易评价方法,比较并筛选一株适合以木薯粉水解液发酵发酵生产燃料酒精的活性干酵母。优化2株酿酒活性干酵母的活化条件、生长温度和生长pH值,以木薯粉水解液为发酵液,在最适生长条件下比较两株酵母的生长曲线、发酵强度、耐糖能力、耐温能力和耐酒精能力。选择酿酒活性干酵母Ⅰ作为木薯粉水解液发酵酵母,该酵母在前36h发酵强度高于1g(/L.h),36h后发酵强度迅速下降;耐糖能力为20%,耐受温度是55℃以及耐酒精浓度是7%。     

Acid, bile, and heat tolerance of free and microencapsulated probiotic bacteria

ABSTRACT:  Eight strains of probiotic bacteria, including Lactobacillus rhamnosus , Bifidobacterium longum, L. salivarius, L. plantarum , L. acidophilus , L. paracasei , B. lactis type Bl-O4, and B. lactis type Bi-07, were studied for their acid, bile, and heat tolerance. Microencapsulation in alginate matrix was used to enhance survival of the bacteria in acid and bile as well as a brief exposure to heat. Free probiotic organisms were used as a control. The acid tolerance of probiotic organisms was tested using HCl in MRS broth over a 2-h incubation period. Bile tolerance was tested using 2 types of bile salts, oxgall and taurocholic acid, over an 8-h incubation period. Heat tolerance was tested by exposing the probiotic organisms to 65 °C for up to 1 h. Results indicated microencapsulated probiotic bacteria survived better ( P < 0.05) than free probiotic bacteria in MRS containing HCl. When free probiotic bacteria were exposed to oxgall, viability was reduced by 6.51-log CFU/mL, whereas only 3.36-log CFU/mL was lost in microencapsulated strains. At 30 min of heat treatment, microencapsulated probiotic bacteria survived with an average loss of only 4.17-log CFU/mL, compared to 6.74-log CFU/mL loss with free probiotic bacteria. However, after 1 h of heating both free and microencapsulated probiotic strains showed similar losses in viability. Overall microencapsulation improved the survival of probiotic bacteria when exposed to acidic conditions, bile salts, and mild heat treatment.     

鼠李糖乳杆菌耐酸及胆盐能力研究

本实验对来源于婴儿粪便鼠李糖乳杆菌进行耐酸、耐胆盐能力表征及模拟胃液和模拟肠液实验。结果获得鼠李糖乳杆菌菌株B在pH1.8条件下,维持2h,活菌数达到107CFU/ml以上;在pH值为2.2以上的培养基中生长良好,而且随着时间的延长菌株的数量增加。在胆盐浓度在0.5%以下的MRS-broth培养基中鼠李糖乳杆菌B菌株维持4h,活菌数均达到108CFU/ml以上,而且随着时间的延长活菌数均有所增加。模拟胃液和模拟肠液实验结果表明该菌株能够有效通过胃环境,并能在肠道中繁殖。结果可见,鼠李糖乳杆菌B具有较强的酸的胆盐的耐受能力,符合微生态制剂和乳酸菌发酵功能食品菌种的要求。     

威宁火腿中微生物的分离鉴定及其耐受特性

以贵州威宁火腿为研究对象,对其中的微生物开展分离鉴定及耐盐、耐亚硝酸盐和耐低温特性研究。结果表明：共分离得到10 株菌,包括4 株马胃葡萄球菌（Staphylococcus equorum）、2 株木糖葡萄球菌（Staphylococcus xylosus）、2 株乳酸片球菌（Pediococcus acidilactici）、1 株变平滑假丝酵母菌（Candida metapsilosis）和1株近平滑假丝酵母菌（Candida parapsilosis）;其中,马胃葡萄球菌WN1、变平滑假丝酵母菌WN9和近平滑假丝酵母菌WN10耐受9% NaCl、150 mg/kg亚硝酸盐和4 ℃低温条件的能力较强。     

Acid and bile tolerance and cholesterol removal ability of lactobacilli strains

Eleven strains of lactobacilli were studied for their acid and bile tolerance. Possible mechanisms of cholesterol removal by strains of lactobacilli were examined. Cholesterol assimilation as determined by the difference in cholesterol content in the medium before and after the incubation period showed that all lactobacilli strains were able to assimilate cholesterol at varying levels ranging from 12.03 to 32.25 microg/mL. Cholesterol removal was associated with growth of cultures. Binding of cholesterol to lactobacilli cells was determined using growing, heat-killed, and resting cells in phosphate buffer. Cholesterol removed by dead and resting cells ranged from 0.79 to 3.82 mg/g of dry weight compared with growing cells, which ranged from 4.53 to 16.03 mg/g of dry weight. Fatty acid methyl esters, as quantified using gas chromatography, showed changes in lipid profiles in cells grown in the presence of cholesterol compared with those grown without cholesterol. Fatty acid profiles, especially of hexadecanoic, octadecanoic, total saturated, and unsaturated acids suggested that cholesterol from the medium was incorporated into the cellular membrane. These findings suggest that strains of lactobacilli could remove cholesterol via various mechanisms and may be promising candidates for use as a dietary adjunct to lower serum cholesterol in vivo.     

几种品牌酵母耐性生长曲线对比研究

选择四种不同品牌的高、低糖酵母,针对其耐盐性和耐糖性生长曲线进行了研究比较.结果表明,无论是高糖或低糖酵母,其盐含量在0～3％时生长情况较好,生长12～16h达到平稳状态;在盐含量≥3％时,四种酵母的耐盐胁迫能力明显减弱.糖含量为0～8％的情况下,适宜四种低糖酵母的生长,且趋势较为接近,都是在16h左右达到一个稳定的阶段.糖含量为0～30％的情况下,高糖酵母的耐糖能力较强,且生长较好.当糖含量≥30％时,其耐糖胁迫能力明显减弱.     

Characterization and adsorption of a Lactobacillus plantarum virulent phage

Bacteriophage infection of lactic acid bacteria is considered one of the biggest worldwide problems in the food industry. Bacteriophages may cause negative effects on the fermentation of various dairy-based products. A virulent bacteriophage was isolated from an abnormal fermentation liquid of Lactobacillus plantarum IMAU10120. The characterization and influence of temperature, pH, divalent cations, and chloramphenicol on the adsorption ability of this phage were evaluated. The results showed that this phage belonged to the Siphoviridae family. It exhibited a burst time of 135 min and a burst size of approximately 215 counts expressed per milliliter per infective center. No significant effect was shown to influence its viability and adsorption at 10 to 37°C. More than 90% of phages exhibited infectivity from pH 5 to 9. Divalent ions and chloramphenicol did not have a significant influence on the adsorption of this phage. The information obtained in this study will enrich the database of lactobacilli virulent phages and provide a basis of information for the control of phages in the food fermentation industry.     

Stability and functionality of freeze-dried probiotic Bifidobacterium cells during storage in juice and milk

We investigated the stability of differently produced (variables being fermentation time, pH during drying, and cryoprotectant) freeze-dried Bifidobacterium animalis subsp. lactis E-2010 (Bb-12) cells in fruit juice and low-fat milk. In addition, the effect of the food matrix on the acid and bile tolerance of the cells was studied. Cells produced in different ways had comparable stability in milk, whereas in juice, sucrose-protected cells survived better than reconstituted skim milk-protected cells. The acid and bile tolerance was better in cells added to milk compared with those in phosphate buffered saline or juice. Despite good culturable stability in milk the acid and bile tolerance of cells decreased during the storage. Apparently, culturable stability data alone do not give an accurate enough prediction of the probiotic functionality in adverse conditions (e.g. survival in acid and bile stress). When choosing a cryoprotectant for a probiotic also the stability in target food applications should be considered.