张国玲,杜伟,刘德华

油脂酵母具有高产油能力,并且所积累油脂的主要成分与植物油脂相似,可作为生物柴油制备的原料.本文对影响酵母油脂合成的关键酶、基因、碳源以及酵母油脂在生物柴油制备中的研究进展进行了综述,认为ATP∶柠檬酸裂解酶和苹果酸酶是酵母油脂合成代谢途径中的关键酶,另外,LRO1、DGA1和ARE基因也被认为同油脂合成有着紧密联系.对酵母油脂用于生物柴油生产的前景进行了展望:利用廉价碳源如甘油、能源作物以及木质纤维素水解液等培养酵母,可有效降低生产成本.在不同催化方法下,酵母油脂均可用于制备生物柴油,这对进一步研究生物柴油的生产应用有着重要意义.     

生物酶法转化酵母油脂合成生物柴油

以一株高产油脂圆红冬孢酵母菌(Rhodosporidum toruloides Y4#)干菌粉为原料,利用酸热法提取了该酵母油脂,并对所得油脂进行了分析. 进一步利用该酵母油脂为原料分别研究了无溶剂体系中三步甲醇法及在叔丁醇介质体系中脂肪酶催化合成生物柴油,发现脂肪酶可以有效转化该酵母油脂制备生物柴油. 在优化反应条件下,生物柴油得率可达90%左右,略低于相同条件下利用精制大豆油合成生物柴油的得率.     

产油酵母利用廉价原料合成油脂的研究进展

利用微生物油脂生产的生物柴油是替代传统化石燃料的绿色可再生能源,产油酵母是极具潜力的产油微生物。然而利用葡萄糖作为碳源成本较高,限制了微生物油脂合成技术的规模化应用,因而寻找廉价原料至关重要。本文基于产油酵母合成油脂的代谢机理,重点介绍了产油酵母利用4种廉价原料,即木质纤维素、粗甘油、有机废水和挥发性脂肪酸合成微生物油脂的研究进展。针对这些来自工农业副产物的廉价原料,分别讨论了其复杂成分对产油酵母生长和油脂合成产生的促进或抑制作用,分析总结了高效预处理、基因工程改造菌体以及微生物共培养等解决方案。通过以上分析,阐明了产油酵母利用廉价原料合成油脂的优点,提出了针对复杂成分和高有机质浓度的抑制性问题的解决方法,理清了未来研究的方向,有助于进一步推动廉价原料用于产油酵母生产油脂及可再生能源技术的发展。     

废旧油脂的精制及酯交换制备生物柴油的研究

采用除杂、脱水、萃取、脱色等操作对废油脂如废煎炸油、地沟油、橡胶籽油进行了精制;对大豆油和精制处理前后的上述废油脂的各种物理化学性质进行了测定和评价;采用共沉淀法制备了Mg/Al类水滑石,450℃焙烧得到复合氧化物并以其催化各种油脂和甲醇合成生物柴油;分别用1H NMR法和仲裁法分析对比酯交换反应的产率。结果表明,大豆油制备生物柴油的产率高达96.9%,而精制后的地沟油、煎炸油和橡胶籽油制备生物柴油的收率分别达38.6%、40.2%和82.0%,可有效降低生物柴油生产成本。尤其是橡胶籽油,有望成为大豆油的替代原料用于生物柴油生产工业。     

3株油脂酵母产油特性的对比研究

微生物油脂可作为生物柴油和其他油脂基化学品的原料,明确产油菌株的产油规律和所产油脂脂肪酸组成是确定其开发潜力的关键。在葡萄糖限氮培养基中对比了3株产油酵母的油脂积累能力,圆红冬孢酵母皮状丝孢酵母禾本红酵母;采用GC-MS分析了3株产油酵母的脂肪酸组成,3株产油酵母油脂均以C16和C18为主,可作为生物柴油的原料油脂。     

高酸值油脂制备生物柴油的方法研究现状

对以高酸值油脂为原料制备生物柴油的降酸工艺进行了研究,综述了酸碱法、两步法、酶催化和超临界法等相关技术,同时介绍了采用高酸值油脂为原料制备生物柴油的研究成果,并对它们各自的优缺点进行了分析,最后展望了高酸值油脂制备生物柴油的发展方向。     

三代生物柴油的制备与研究进展

综述了三代生物柴油的制备和工艺条件,包括第一代生物柴油制备方法:酸碱催化法、酶催化法和超临界法;第二代生物柴油制备方法:掺炼法、加氢直接脱氧法和加氢脱氧异构法;第三代生物柴油制备方法:微生物油脂法、生物质气化合成法。     

中国生物柴油技术开发及应用进展

综述了目前我国制备生物柴油的主要廉价原料资源即基因改造植物油脂、废弃油脂和微生物油脂,并介绍了现阶段我国生物柴油的酯交换技术开发和应用现状,而且对生物柴油的发展前景进行了展望,指出必须走出一条符合我国国情的生物柴油发展道路。     

游离脂肪酶NS81006催化含酸油脂制备生物柴油的应用研究

与固定化脂肪酶相比,游离脂肪酶具有反应速率快、成本较低的优势,成为制备生物柴油新的研究方向。前期研究结果表明,游离脂肪酶NS81006可以高效催化大豆油甲醇解制备生物柴油,进一步研究其对含酸油脂的催化,对于促进游离脂肪酶在生物柴油领域中的应用具有重要意义。本文系统研究了甲醇添加策略对游离脂肪酶NS81006催化油酸制备生物柴油的影响,进而考察了NS81006催化模拟酸化油以及实际含酸油脂制备生物柴油的转化情况。研究表明,在优化的甲醇添加策略下,游离脂肪酶NS81006可有效催化油酸、不同含酸量(0~100%,基于总质量)模拟酸化油以及实际含酸油脂进行生物柴油的制备;离心分离可有效实现NS81006的回复使用,连续回用5个批次,游离脂肪酶活性未出现明显下降。     

固体酸催化制备生物柴油研究进展

生物柴油是一种具有优良燃烧性能的石化替代燃料,开发可用于通过酯交换反应制备生物柴油的固体酸催化剂已成为国内外研究热点。本文综述了固体酸在催化合成生物柴油方面的研究进展,介绍了以纳米材料、磁性材料作为载体以及掺杂稀土元素等固体酸改性方法,并对固体酸催化制备生物柴油的发展前景进行了展望。     

Production of lipid from N‐acetylglucosamine by Cryptococcus curvatus

N‐Acetylglucosamine (GlcNAc), the monomeric constituent of chitin, is rarely used as a carbon source for fermentation technology. In this study, we demonstrate that the oleaginous yeast Cryptococcus curvatus ATCC 20509 can produce intracellular lipid during the cultivation process and total lipid content can reach 54% on a GlcNAc‐based medium. Culture of C. curvatus under various conditions indicated that lipid accumulation also occurred at a relatively broad range of temperatures as well as relatively high initial GlcNAc concentrations. Fatty acid analysis indicated that the product was rich in palmitic acid, stearic acid, and oleic acid, closely resembling the composition of palm oil. More importantly, the lipid sample produced at 22 °C had a total saturated fatty acid content of 54.2 wt%, suggesting that it may be explored as cocoa‐butter equivalent. Our data suggested that GlcNAc could be used as a feedstock for industrial biotechnology and that C. curvatus ATCC 20509 is a strain capable of accumulating high intracellular lipid using this nitrogen‐rich renewable material. Practical applications: Microbial lipid is a versatile material, especially for biodiesel production. Stable and abundant renewable raw substrates remain to be explored for large‐scale production of microbial lipid. The present work reports lipid production using N‐acetylglucosamine (GlcNAc) by the oleaginous yeast Cryptococcus curvatus ATCC 20509 to yield up to 54% intracellular lipid content. More significantly, the lipid sample produced at 22 °C had a total saturated fatty acid content of 54.2 wt%, suggesting that it may be explored as cocoa‐butter equivalent. Our technology provides the opportunity to effectively convert GlcNAc, available from one of the most abundant renewable materials chitin, into lipid. This procedure should prove valuable in terms of renewable energy production as well as environmental pollution control.     

基于消耗碳氮比进行底物分配和油脂得率预测

微生物油脂作为胞内代谢产物,其脂肪酸组成和植物油类似,是生物柴油和油脂工业理想替代原料。为了进一步了解底物中碳氮比对油脂合成的影响,通过恒化培养的方法,研究了圆红冬孢酵母在不同稀释率条件下,消耗碳氮比和底物在油脂和非油生物量之间的分配以及和油脂得率之间的关系。通过碳、氢、氧和氮的化学反应计量学分析,并根据不同稀释率稳态时所消耗C/N比,构建了底物碳在油脂和非油生物量之间的分配模型。利用实验数据确定了模型参数:最大油脂碳得率Y_L^max为0.51 mol C·(mol C)^-1,最大菌体碳得率Y_L^max为0.52 mol C·(mol C)^-1,促使油脂合成的临界C/N比为12.1 mol C·(mol C)^-1。利用该分配模型预测不同消耗C/N比的油脂得率,预测值为实验值的 95.2%～116.7%,表明模型可靠性较好。     

Advances in cultivation and processing techniques for microalgal biodiesel: A review

The key technologies for producing microalgal biodiesel include microalgae screening, economical cultivation, and efficient methods in lipid extraction and conversion. Recent advances in microalgae cultivation, lipid extraction, and biodiesel preparation are reviewed in this work, with emphasis on photosynthetic metabolisms, separation efficiency and catalytic kinetics. The mutual exclusion between lipid accumulation and fast growth limits total lipid productivity, while only triglycerides in neutral lipids are converted to biodiesel through transesterification. The hurdles in large scale culture and low neural lipids yield are discussed, as well as the relationship between high unsaturation and fuel properties. This review aims to provide technical information to guide strain screening and lipid conversion for microalgal biodiesel industry.     

Lipids of oleaginous yeasts. Part II: Technology and potential applications

The process of lipid accumulation in the oleaginous yeasts cultivated in various fermentation configurations when either sugars and related compounds or hydrophobic substances are used as substrates is presented and kinetic models describing both de novo and ex novo lipid accumulation are analyzed. Technological aspects related with single cell oil (SCO) produced by oleaginous yeasts are depicted. The influence of culture parameters upon lipid production process is presented. Lipid production has been studied in batch, fed‐batch, and continuous cultivation systems using yeasts belonging to the species Lipomyces starkeyi, Rhodosporidium toruloides, Apiotrichum curvatum, Candida curvata, Cryptococcus curvatus, Trichosporon fermentans, and Yarrowia lipolytica. The potentiality of yeasts to produce SCO as starting material of 2nd generation biodiesel is indicated and discussed. Of significant importance is also the utilization of yeast lipids as substitutes of high added value exotic fats (e.g., cocoa butter). Lipid produced by the various yeasts presents, in general, similar composition with that of common vegetable oils being composed of unsaturated fatty acids, whereas cocoa butter is principally composed of saturated fatty acids, consequently the various strategies that are followed in order to increase the cellular saturated fatty acid content of the yeast lipid are presented and comprehensively discussed.     

Enhanced microbial oil production by activated sludge microorganisms via co‐fermentation of glucose and xylose

The co‐fermentation of glucose and xylose by activated sludge microorganisms for the production of microbial oils for use as biodiesel feedstock was investigated. Various carbon sources at initial concentration of 60 g/L and C:N ratio 70:1 were investigated: xylose, glucose, and 2:1 and 1:2 (by mass) glucose/xylose mixtures. Oil accumulation ranged between 12 to 22% CDW, the highest of which was obtained when xylose was the sole substrate used. Kinetic modeling of the fermentation data showed that specific growth and oil accumulation rates were similar in all substrate types and the lipid coefficient ranged from 0.02 to 0.06 g/g of sugar consumed. The fatty acid methyl ester yield and composition of the lipids showed their suitability for conversion to biodiesel. Based on the results, lignocellulose sugars could be used as fermentation substrates by activated sludge microorganisms for enhancing the oil content of sewage sludge for its use as a sustainable biofuel feedstock source. © 2013 American Institute of Chemical Engineers AIChE J, 59: 4036–4044, 2013     

New Biofuel Alternatives: Integrating Waste Management and Single Cell Oil Production

Concerns about greenhouse gas emissions have increased research efforts into alternatives in bio-based processes. With regard to transport fuel, bioethanol and biodiesel are still the main biofuels used. It is expected that future production of these biofuels will be based on processes using either non-food competing biomasses, or characterised by low CO₂ emissions. Many microorganisms, such as microalgae, yeast, bacteria and fungi, have the ability to accumulate oils under special culture conditions. Microbial oils might become one of the potential feed-stocks for biodiesel production in the near future. The use of these oils is currently under extensive research in order to reduce production costs associated with the fermentation process, which is a crucial factor to increase economic feasibility. An important way to reduce processing costs is the use of wastes as carbon sources. The aim of the present review is to describe the main aspects related to the use of different oleaginous microorganisms for lipid production and their performance when using bio-wastes. The possibilities for combining hydrogen (H₂) and lipid production are also explored in an attempt for improving the economic feasibility of the process.     

Anthracene Biodegradation by Oleaginous Rhodococcus opacus for Biodiesel Production and Its Characterization

This study examined biodegradation of anthracene, a model low molecular weight polycyclic aromatic hydrocarbon (PAH) by oleaginous Rhodococcus opacus for biodiesel production. Specific biomass growth rate (µ) in the range of 0.0075–0.0185 h^?1 could be attained over the initial anthracene concentration (50–500 mg L^?1), along with 68–70.6% (w/w) lipid accumulation. 10% (v/v) inoculum size showed more positive effect than 5% (v/v) inoculum size on both anthracene biodegradation efficiency and lipid accumulation by R. opacus. ¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy of the bacterial lipids revealed 82.25% saturated fatty acids content. Furthermore, the transesterified bacterial lipids predominantly consisted of methyl palmitate (32.4%) and methyl stearate (25.9%) as the major fatty acid methyl esters (FAMEs). Overall, this study revealed a very good potential of the bacterium for the production of biodiesel from PAH-containing wastewater.     

Microbial lipid production as biodiesel feedstock from N‐acetylglucosamine by oleaginous microorganisms

BACKGROUND: The byproducts from shrimp processing are heads and shells which contain a wealth of carbon and could be converted into oils via oleaginous microorganisms. The objective of this investigation was to determine the feasibility of using oleaginous microorganisms to convert N‐acetylglucosamine (GlcNAc), the major carbohydrate of the hydrolysate of shrimp processing waste, to triacylglycerols as a biodiesel feedstock. RESULTS: Screening experiments were conducted among Rhodotorula glutinis, Rhodococcus opacus and Cryptococcus curvatus using GlcNAc as sole carbon and energy source at 30 °C. All three microorganisms were found to grow well on GlcNAc, but the lipid contents in the cells were consistently low (lower than 5%) in the growth phase. However, lipid accumulation by C. curvatus was greatly enhanced upon entering the death phase in the absence of GlcNAc and the lipid content increased to 28.4% at 167.7 h. This indicated that C. curvatus was the optimal tested microorganism for the production of microbial oils from GlcNAc. Phosphate was further evaluated on the growth and lipid production by C. curvatus from GlcNAc. Results indicated that the yields of both biomass during growth phase and lipids at death phase increased with the increase of the ratio of C to P. But the fatty acid profiles of the accumulated lipids did not change significantly. CONCLUSION: Results indicated that shrimp processing waste could be utilized to produce oils as a biodiesel feedstock. The results could be applied to maximize production of oils from shrimp processing waste. Copyright © 2011 Society of Chemical Industry     

利用可再生油料资源发展生物炼油化工厂

石油、天然气、煤炭资源终将枯竭,利用可再生资源生产燃料和化工产品正在兴起.利用植物油与甲醇酯交换,可以制备脂肪酸甲酯.脂肪酸甲酯是一种优质清洁燃料,即生物柴油;同时,脂肪酸甲酯及其联产的甘油进一步精制和深加工可以生产许多化工产品.本文从生物柴油的国内外发展概况出发,结合国内的现状,对我国发展生物柴油的影响因素进行了分析,认为原料油的价格和供应量是关键;同样,也对生产化工产品作了相应的探讨,认为这是增加利润的关键.最后形成利用可再生油料资源发展炼油化工厂的设想.     

Enzymatic transesterification for production of biodiesel using yeast lipases: An overview

Biodiesel has provided an eco-friendly solution to fuel crisis, as it is renewable, biodegradable and a non-toxic fuel that can be easily produced through enzymatic transesterification of vegetable oils and animal fats. Enzymatic production of biodiesel has many advantages over the conventional methods as high yields can be obtained at low reaction temperatures with easy recovery of glycerol. Microbial lipases are powerful biocatalysts for industrial applications including biodiesel production at lower costs due to its potential in hydrolyzing waste industrial materials. Among them, lipases from yeasts, Candida antarctica, Candida rugosa, Cryptococcus sp., Trichosporon asahii and Yarrowia lipolytica are known to catalyze such reactions. Moreover, stepwise addition of methanol in a three step, two step and single step reactions have been developed using yeast lipases to minimize the inhibitory effects of methanol. The latest trend in biodiesel production is the use of whole-cell as biocatalysts, since the process requires no downstream processing of the enzyme. Synthesis of value added products from the byproduct glycerol further reduces the production cost of biodiesel. This review aims at compiling the information on various yeast lipase catalyzed transesterification reactions for greener production of biodiesel.