圆红冬孢酵母两阶段培养法生产微生物油脂

为缩短发酵时间,减少原料消耗,采用细胞增殖和油脂积累分离的两阶段模式,培养圆红冬孢酵母Rhodosporidium toruloides AS 2.1389生产微生物油脂。结果表明,细胞增殖阶段获得的R.toruloides AS 2.1389细胞,重悬接种在葡萄糖溶液中,可快速积累油脂,菌体油脂含量超过自身干重的55%。增殖阶段细胞的菌龄越高,产油能力越强。油脂积累阶段使用高浓度葡萄糖溶液或未灭菌的葡萄糖溶液,油脂合成可以有效进行。油脂中脂肪酸成分以含16和18个碳原子的长链脂肪酸为主,可作为制备生物柴油的新型原料。     

发酵性丝孢酵母HWZ004利用水稻秸秆水解液发酵产油脂

为高效利用水稻秸秆中的纤维素和半纤维素产油脂,采用稀酸预处理和酶水解两步法对水稻秸秆进行水解,然后以水解液为碳源,培养发酵性丝孢酵母Trichosporon fermentans HWZ004产微生物油脂。结果表明,经简单overliming法脱毒后水稻秸秆水解液中乙酸、糠醛和5-羟甲基糠醛的浓度分别为0.4 g/L、0.1 g/L和0.05 g/L。只需添加少量氮源和微量CuSO4?5H2O,该水解液即可满足T. fermentans HWZ004发酵产油脂的要求。发酵最适接种量、初始pH和温度分别是5.0％、7.0和25 ℃。T. fermentans HWZ004在优化条件下培养7 d的生物量、油脂含量和油脂产量分别是26.4 g/L,52.2％和13.8 g/L;油脂得率系数为17.0,大大高于驯化前菌株T. fermentans CICC 1368在脱毒水稻秸秆半纤维素水解液中的对应值 (11.9)。所产油脂的脂肪酸组成与植物油相似,不饱和脂肪酸含量达70％以上,宜作为生物柴油的生产原料。     

斯达氏油脂酵母利用混合糖发酵产油脂

研究了斯达氏油脂酵母Lipomyces starkeyi2#利用葡萄糖-木糖混合糖为碳源生长和油脂积累特性。L.star-keyi2#利用70 g/L葡萄糖和70 g/L木糖作为碳源在30℃下摇瓶发酵96 h,糖利用率均达90%以上,菌体生物量分别为14.1 g/L和13.1 g/L,油脂质量分数分别为55.7%和52.6%。相同条件下该菌株利用混合糖(葡萄糖46 g/L,木糖24 g/L)为碳源时总糖利用率、生物量和油脂质量分数分别为75.1%,15.0 g/L和40.0%。借助于P lackett-Burm an设计法和单因子实验法对培养条件进行了优化,结果表明发酵96 h混合糖利用率可达到97.3%,发酵120 h后混合糖利用率、生物量和菌体油脂质量分数分别达99.5%、19.0 g/L和52.6%。生物量得率和油脂得率分别达到27%和14%。     

粘红酵母在味精废水中发酵生产油脂

对粘红酵母菌株进行驯化得到一株优良菌株Rh8,利用其在味精废水中发酵以去除废水中的化学需氧量(COD)并生产油脂;考察废水pH以及添加葡萄糖母液、营养因子等对菌株Rh8在味精废水中生长、产油和COD去除效果的影响,发现将废水稀释4倍、调节pH至5.5时,菌株可以较好地生长;而添加废葡萄糖母液、酵母粉、KH2PO4、MgSO4、MnSO4均能够促进菌体的生长、产油和废水中的COD去除,在250 mL摇瓶中,生物量最高可达15.6 g/L,干菌体中油脂质量分数达到29.61%,COD去除率达到45.1%。     

圆红冬孢酵母发酵菊芋块茎产油脂的研究

研究了圆红冬孢酵母Y4发酵菊芋块茎,菊芋品种及其处理方法对发酵产油的影响。结果表明,菊芋浸提汁、酸水解液或菊芋浆均可直接被圆红冬孢酵母Y4利用,发酵积累油脂,但白皮菊芋比紫皮菊芋更有利于油脂发酵。发酵菊芋浸提汁或酸水解液时,无需添加外源营养物,干菌体油脂含量可达到40%（w/w）;发酵菊芋浆时,白皮菊芋转化率达到12.1 g油/100 g去皮干菊芋。菊芋油脂发酵产品主要以16碳和18碳系脂肪酸为主,与常规植物油的脂肪酸组成相似,可作为制备生物柴油的新型替代原料。     

β-1,3-葡甘露聚糖酶辅助提取圆红冬孢酵母油脂

为考察β-1,3-葡甘露聚糖酶辅助提取圆红冬孢酵母Rhodosporidium toruloides油脂工艺的应用潜力,以R. toruloides Y4发酵醪液为原料,乙酸乙酯为溶剂,在10 L规模下研究了酶解和萃取条件对油脂提取率的影响,并进行了初步物料平衡分析。结果表明：酶处理0.5 h,油脂提取率可达71.1%,酶解效率与10 mL小体系所取得的结果相当。通过多次萃取,有效缓解了乳化现象对油脂提取的影响。经过3次萃取,油脂提取率、溶剂回收率和总物料回收率分别达92.9%、87.0%和94.2%。酶辅助提取油脂工艺可直接利用油脂发酵醪液,提取率高,对设备要求低,具有很好的工业化应用潜力。     

皮状丝孢酵母B3利用木薯淀粉发酵生产微生物油脂

对皮状丝孢酵母B3以木薯淀粉水解液为碳源发酵生产微生物油脂培养条件进行了优化,并在2 L发酵罐中对菌体生长和油脂积累进行了考察。摇瓶实验表明,木薯淀粉水解液的浓度高于90 g/L时不利于菌体的生长和油脂积累,皮状丝孢酵母B3发酵生产微生物油脂的最适氮源及其浓度、最适C/N比和pH分别为酵母提取物3.0 g/L、116、6.0,在此条件下培养144 h菌体生物量、油脂产量和油脂含量分别达到15.2 g/L、6.22 g/L和40.9％;在2 L发酵罐中分批发酵44 h后菌体生物量、油脂产量和油脂含量分别达28.7 g/L、12.27 g/L和42.8％。以皮状丝孢酵母B3所产油脂制备生物柴油,其主要组成包括棕榈酸甲酯、硬脂酸甲酯、油酸甲酯、亚油酸甲酯等,且理化特性符合相关国家标准,可作为一种有潜力的化石燃料替代品。     

利用蔗渣半纤维素水解液产油酵母的筛选、鉴定及发酵实验

对已分离到的产油酵母通过木糖摇瓶发酵,从中筛选到2株H2-1和J2-2利用蔗渣半纤维素水解液高产油脂酵母,其利用蔗渣半纤维素水解液油脂得率系数分别为13.49和10.28,均显示出对蔗渣半纤维素水解液的高转化率.根据常规生理生化特征和26S rDNA序列分析结果,确认H2-1为小红酵母(Rhodotorula minuta),J2-2为粘红酵母(Rhodotorula glutinis).     

不同营养元素限制对圆红冬胞酵母油脂生产的影响

以产油酵母圆红冬胞酵母（Rhodosporidium toruloides）作为研究对象,系统地研究了氮、磷、硫限制对其油脂积累的影响,并在3L生物反应器上考察了R.toruloides在C/P摩尔比为1 133.3时初始葡萄糖浓度对油脂生产的影响。结果表明：氮、磷、硫中任意一种营养元素受限,均能促使R.toruloides在胞内积累高于自身干重60%的油脂;通过改变培养基的组成,可以调节油脂中脂肪酸的构成,使油脂中饱和脂肪酸比例高于70%或不饱和脂肪酸比例高于60%。就油脂生产强度及转化效率而言,磷限制优于氮限制或硫限制。当C/P摩尔比相同时,初始葡萄糖浓度越低越有利于油脂生产。对采用不同原料生产微生物油脂的技术有一定指导意义。     

Recycling biodiesel-derived glycerol by the oleaginous yeast Rhodosporidium toruloides Y4 through the two-stage lipid production process

Direct utilization of crude glycerol, a major byproduct in biodiesel industry, becomes imperative, because its production has outpaced the demand recently. We demonstrated that the oleaginous yeast Rhodosporidium toruloides Y4 had a great capacity to convert glycerol into lipids with high yield using the two-stage production process. Significantly higher cell mass and lipid yield were observed when the media were made with synthetic crude glycerol than pure glycerol. The process achieved a lipid yield of 0.22 g g⁻¹ glycerol, which was comparable with the lipid yield using glucose as the substrate. Lipid samples showed similar fatty acid compositional profiles to those of vegetable oils, suggesting that such microbial lipids were potential feedstock for biodiesel production. Our data provided an attractive route to integrate biodiesel production with microbial lipid technology for better resource efficiency and economical viability.     

Lipid production by Lipomyces starkeyi cells in glucose solution without auxiliary nutrients

Two-stage fermentation process was used for lipid production by Lipomyces starkeyi AS 2.1560 in glucose solution without auxiliary nutrients. In the first stage, cells were cultivated in a nutrient-rich medium for propagation. In the second stage, cells were resuspended in glucose solution to achieve high cellular lipid contents. The effects of the inocula age, cell density and initial glucose concentration on lipid production were briefly studied. When high cell density fermentation was performed in a 7-L stirred-tank bioreactor for 40 h using non-sterile glucose solution as carbon source, the biomass, lipid and lipid content reached 104.6 g/L, 67.9 g/L and 64.9%, respectively. More significantly, lipid productivity reached 2.0 g/L h during the initial 16 h-period and 1.6 g/L h for the entire culture. Our results demonstrated that cell propagation and lipid accumulation processes can be spatially separated, allowing further optimization to improve both processes. The two-stage fermentation method should have a great potential to develop more efficient processes to convert renewable materials into biofuel and related products.     

Production of microbial lipid by Cryptococcus curvatus on rice straw hydrolysates

Microbial biolipids/biodiesels derived from volatile fatty acids (VFAs) can be a valuable alternative to plant oils if optimum fermentation conditions are determined. VFAs were used for cell mass and microbial lipid production by Cryptococcus curvatus. The lipid content in the cells increased up to 48% and 28% in batch cultures with the use of 20 g/L glucose and 6 g/L of VFAs as the carbon source, respectively. In this study, C. curvatus used VFAs as a carbon source via anaerobic digestion of rice straw hydrolysates. VFAs produced from rice straw resulted in yield of 0.43 g VFAs/g substrate and 40% higher specific growth rate(0.305 h⁻¹) than synthetic VFAs. The highest fatty acid composition observed was C18:1, was obtained using glucose and VFAs as the carbon source to yield a cetane number of 56–59, which is suitable for biodiesel production. The cost of microbial lipids was estimated to be 0.30–1.15 USD/L given 0–150 USD/ton of VFAs cost for a yield of 0.17 g/g of lipids. Thus, VFAs can be a suitable carbon source for economical biodiesel production.     

油脂酵母发酵木糖生产微生物油脂

目的用斯达油脂酵母(Lipomyces starkeyi)作为发酵菌株,以纯木糖溶液为油脂发酵原料,对L.starkeyi利用木糖积累油脂进行系统研究。方法 L.starkeyi于斜面培养基中活化后,接种于YPD液体培养基,于30℃、200 r/min摇床培养。在摇瓶中培养一段时间后,测定发酵液细胞浓度,离心发酵液收集细胞。将离心后得到的菌体加入木糖溶液重悬,并转接于含50 mL木糖溶液的250 mL摇瓶中进行发酵生产。结果相比一阶段法,两阶段发酵方法可以在更短的时间内达到较高的油脂含量,油脂含量能够达到细胞自身干重的60%以上。实验发现高菌龄酵母产油速度更快;并且初始木糖浓度高达120 g/L时,酵母细胞仍然能够高效合成油脂。结论 L.starkeyi能够有效利用木糖进行发酵产生油脂,是以木质纤维素为原料生产微生物油脂的优良菌种。