生物柴油植物黑皂树开发利用研究

能源短缺和环境污染是当前人类社会所面临的巨大挑战,生物柴油的推广和应用是现阶段解决替代燃油的重要途径之一。黑皂树种子油是很好的生物柴油原料,目前在许多热带地区大力种植黑皂树。我国华南、西南地区的一些干旱荒地可以种植黑皂树。我们对黑皂树油在柴油机上的应用研究证明:黑皂树油在气温35℃条件下,可以直接在各种柴油机上起动和应用;黑皂树油中掺入20%的轻柴油,其黏度大大降低,冷机起动性能和热效率等均与轻柴油相当。因此,黑皂树油的应用比目前国外使用的生物柴油更经济,是一种很有发展前途的能源植物。     

Biodiesel production from Jatropha oil by catalytic and non-catalytic approaches: an overview

Biodiesel (fatty acids alkyl esters) is a promising alternative fuel to replace petroleum-based diesel that is obtained from renewable sources such as vegetable oil, animal fat and waste cooking oil. Vegetable oils are more suitable source for biodiesel production compared to animal fats and waste cooking since they are renewable in nature. However, there is a concern that biodiesel production from vegetable oil would disturb the food market. Oil from Jatropha curcas is an acceptable choice for biodiesel production because it is non-edible and can be easily grown in a harsh environment. Moreover, alkyl esters of jatropha oil meet the standard of biodiesel in many countries. Thus, the present paper provides a review on the transesterification methods for biodiesel production using jatropha oil as feedstock.     

生物柴油植物膏桐高油高产品种的培育(英文)

在2005-2009年期间,利用从中国南部和西南部收集的80个不同的膏桐地理种源,在西双版纳建立膏桐种质资源圃30亩;在圃内开展其生物学特性和农艺性状的同时,对其中表现良好的6个地理种源做了进一步选择和培育。结果表明:80个种源的2年生时平均直径、树高和树冠分别为7.6cm、167cm和114cm,种子千粒重为0.676(0.477～0.879)kg;在这些种源中,根据单株产量和种子含油率的情况,对6个表现较好的种源开展了小规模试验,2年生时平均单株产量0.34kg,3年生时达1.38kg;在试验中,有一个种源表现尤为突出,自挂果以来种子产量逐年增加,二、三、四年生林分每公顷干种子产量分别为964.3kg、2000.6kg、2858.7kg,种子含油率40%～42%。另外,还利用野外发现的膏桐突变体成功地育出了膏桐新品种。以此为父本与其他种源杂交产生的后代,种子含油率较对照提高6个百分点,杂种种子含油率达41.2%。     

三种麻疯树种子油脂理化特性比较

以海南近海岸陆地上正常生长、结实的3个不同基因型麻疯树（南油1、2和3号）种子为材料,分析比较3种种子及其种子油的理化特性.结果表明：南油1、2和3号麻疯树种子长、宽、厚度和表面积差异均不显著,但南油2号种子千粒重显著大于1、3号、不完善粒率显著小于1、3号,其种子饱满度好,结实性状优.南油3号种子种仁含油量显著高于1、2号,而1、2号间无显著差异.3种麻疯树种子油的过氧化值、折射率和皂化值无显著差异,而南油2号的酸值显著低于1、3号;三者碘值皆低于100,且南油2号的碘值显著小于1、3号.3种麻疯树油脂肪酸成分主要是油酸、棕榈酸、亚油酸、硬脂酸和十七碳酸,且皆以不饱和脂肪酸为主,南油2号的饱和脂肪酸含量相对较高.与南油1号相比, 南油2、3号的应用潜力相对大.     

Life cycle assessment of biofuels from Jatropha curcas in West Africa: a field study

In recent years, liquid biofuels for transport have benefited from significant political support due to their potential role in curbing climate change and reducing our dependence on fossil fuels. They may also participate to rural development by providing new markets for agricultural production. However, the growth of energy crops has raised concerns due to their high consumption of conventional fuels, fertilizers and pesticides, their impacts on ecosystems and their competition for arable land with food crops. Low-input species such as Jatropha curcas , a perennial, inedible crop well adapted to semiarid regions, has received much interest as a new alternative for biofuel production, minimizing adverse effects on the environment and food supply. Here, we used life-cycle assessment to quantify the benefits of J. curcas biofuel production in West Africa in terms of greenhouse gas emissions and fossil energy use, compared with fossil diesel fuel and other biofuels. Biodiesel from J. curcas has a much higher performance than current biofuels, relative to oil-derived diesel fuels. Under West Africa conditions, J. curcas biodiesel allows a 72% saving in greenhouse gas emissions compared with conventional diesel fuel, and its energy yield (the ratio of biodiesel energy output to fossil energy input) is 4.7. J. curcas production studied is eco-compatible for the impacts under consideration and fits into the context of sustainable development.     

Transformation of Jatropha curcas L. for production of larger seeds and increased amount of biodiesel

The development of green energy is important to mitigate global warming. Jatropha (Jatropha curcas L.) is a promising candidate for the production of alternative biofuel, which could reduce the burden on the Earth’s resources. Jatropha seeds contain a large quantity of lipids that can be used to produce biofuel, and the rest of the plant has many other uses. Currently, techniques for plant genetic transformation are extensively employed to study, create, and improve the specific characteristics of the target plant. Successful transformation involves the alteration of plants and their genetic materials. The aim of this study was to generate Jatropha plants that can support biofuel production by increasing their seed size using genes found via the rice FOX-hunting system. The present study improved previous protocols, enabling the production of transgenic Jatropha in two steps: the first step involved using auxins and dark incubation to promote root formation in excised shoots and the second step involved delaying the timing of antibiotic selection in the cultivation medium. Transgenic plants were subjected to PCR analysis; the transferred gene expression was confirmed via RT-PCR and the ploidy level was investigated. The results suggest that the genes associated with larger seed size in Arabidopsis thaliana, which were found using the rice FOX-hunting system, produce larger seeds in Jatropha.     

麻风果油为原料的生物柴油生产全过程能值分析

生物柴油作为石油的替代能源之一,已在我国开始了产业化应用。采用能值分析方法对以麻风果油为原料的生物柴油生产全过程,包括麻风树的种植、麻风果实的运输、麻风果油的榨取、果油酯化4个过程以及废水的处理进行了评价,从可更新资源、不可更新资源和购买资源入手构建了能值评价指标体系,并与大豆油为原料的生物柴油、小麦为原料的生物乙醇进行了比较。结果表明麻风果油生物柴油的能值转换率是1.67×1013sej/kg,能值产出率是1.85,环境负载率是6.84,可持续发展指数是0.271。比较得出,麻风果油能值转换率是大豆油的3倍,是生物乙醇的2倍;相同经济能值的投入,麻风果油能值产出率最高,但对环境的依赖程度也更强;可持续发展指数三者相差不大。原料的非食用性方面,与大豆生物柴油、生物乙醇相比,麻风果油有很大的优势,具有发展潜力。     

Rapid analysis of Jatropha curcas gene functions by virus-induced gene silencing

Jatropha curcas L. is a small, woody tree of the Euphorbiaceae family. This plant can grow on marginal land in the tropical and subtropical regions and produces seeds containing up to 30% oil. Several Asian countries have selected Jatropha for large scale planting as a biodiesel feedstock. Nevertheless, Jatropha also possesses several undesirable traits that may limit its wide adoption. An improved understanding of plant development and the regulation of fatty acid (FA) and triacylglyceride biosynthesis in Jatropha is particularly facilitative for the development of elite crops. Here, we show that a tobacco rattle virus (TRV) vector can trigger virus-induced gene silencing (VIGS) in Jatropha. Our optimized method produced robust and reliable gene silencing in plants agroinoculated with recombinant TRV harbouring Jatropha gene sequences. We used VIGS to investigate possible functions of 13 Jatropha genes of several functional categories, including FA biosynthesis, developmental regulation and toxin biosynthesis, etc. Based on the effects of VIGS on the FA composition of newly emerged leaves, we determined the function of several genes implicated in FA biosynthesis. Moreover, VIGS was able to discriminate independent functions of related gene family members. Our results show that VIGS can be used for high-throughput screening of Jatropha genes whose functions can be assayed in leaves.     

研究开发燃料油植物生产生物柴油的几个策略

能源短缺和环境污染是目前人类社会所面临的巨大挑战,而生物柴油的应用和推广正是现阶段解决能源替代问题的较佳手段。现今国外生物柴油产业发展十分迅速,产量逐年增长,而我国的生物柴油产业才刚刚起步。本文介绍了极具潜力的5种木本油料植物麻疯树（Jatropha curcas）、光皮树（Cornus wilsoniana）、文冠果（Xanthoceras sorbifolia）、黄连木（Pistacia chinensis）和欧李（Cerasus humilis）和1种野生草本油料植物海篷子（Salicornia bigelivii）,进而提出运用转基因技术提高燃料油植物种子含油量的优势,归纳总结了生产生物柴油的4种不同工艺。最后建议政府应对燃料油植物种植和生产加工产业实施补贴和免税等扶植政策。本文对我国生物质能源产业的发展提供了有价值的实施策略,具有重要的借鉴和参考价值。     

麻疯树脂酶全长基因克隆、表达及其蛋白质结构预测

脂酶(Lipase,EC3.1.1.3)是普遍应用于皮革、饲料及生物柴油工业的工业酶制剂,具有广泛的应用价值。目前对植物来源的脂酶研究较少。本研究用在生物柴油中具有应用前景的油料植物——麻疯树(Jatrophacurcas)作为研究对象,克隆了该物种的脂酶基因(JcLIP)。通过多序列比对并结合物种的亲缘关系设计了具有较高特异性的简并引物,通过使用RT-PCR和RACE技术,最终获得了麻疯树脂酶基因的全长序列并成功地在大肠杆菌中表达,酶活测定结果表明,麻疯树脂酶在大肠杆菌中表达在包涵体中,但是能产生具有活力的蛋白质,酶活约为0.8U.mL-1。结构预测和比较表明,JcLIP蛋白质具有脂酶的结构核心和催化活性中心,而在非核心区具有较毛霉脂酶更多的插入和随机卷曲,这可能是决定二者之间酶活差异的重要原因。     

麻疯树种子的研究进展

麻疯树（Jatropha curcas L．）为大戟科（Euphorbiaceae）麻疯树属半肉质小乔木或大灌木,具有很强的抗旱、耐贫瘠的特性。麻疯树的根、树皮、叶和种子均可人药。种子中主要含有脂肪类物质、蛋白质和萜类物质,其毒素为麻疯树毒蛋白和种子油。种仁中的含油量约为50％,可作为理想的生物柴油;毒蛋白、种子油及其他种子提取物可作为生物农药。关于麻疯树种子的发育、脱水行为及其调控研究较少。麻疯树是二种具有重要经济价值的战略资源。     

麻疯树皮的化学成分研究

从麻疯树(Jatrophac curcus L.)的树皮中分离得到12个化合物,经理化常数和波谱鉴定(IR,1H NMR,13C NMR,EI-MS)分别为二十六酸甲酯(1),β-谷甾醇(2),蒲公英萜醇(3),蒲公英甾醇(4),伪蒲公英甾醇(5),curcusones A(6),curcusones B(7),jatropholone A(8),jatropholone B(9),3,3′,4-三甲氧基鞣花酸(10),胡萝卜甙(11),蔗糖(12)。其中化合物1,4,5,10,12为首次从该植物中分离得到。     

从麻疯树上胚轴外植体再生植株

以麻疯树上胚轴为实验材料在MS添加IBA和BA的培养基上进行离体培养实验.结果表明,在IBA O.1 mg/L与BA 0.2～0.7 mg/L组合的条件下,不定芽从上胚轴外植体的表面直接被诱导分化,其中以在MS IBA 0.1 mg/L BA 0.5 mg/L上的诱导率最高.从愈伤组织来源的植株再生需要IBA 0.5 mg/L与BA 0.1 mg/L、IBA 0.5 mg/L与BA0.2mg/L以及IBA1.0mg/L与BA0.5mg/L的激素组合,其分化的最佳培养基是MS IBA1.0 mg/L BA0.5 mg/L.生长健壮的不定芽和再生植株能在无激素的MS基本培养基上生根.发育良好的再生苗可成功地转移到温室栽培而没有可见的变异.     

Fatty Acid Diversity is Not Associated with Neutral Genetic Diversity in Native Populations of the Biodiesel Plant Jatropha curcas L.

Jatropha curcas L. (Euphorbiaceae) is a shrub native to Mexico and Central America, which produces seeds with a high oil content that can be converted to biodiesel. The genetic diversity of this plant has been widely studied, but it is not known whether the diversity of the seed oil chemical composition correlates with neutral genetic diversity. The total seed oil content, the diversity of profiles of fatty acids and phorbol esters were quantified, also, the genetic diversity obtained from simple sequence repeats was analyzed in native populations of J. curcas in Mexico. Using the fatty acids profiles, a discriminant analysis recognized three groups of individuals according to geographical origin. Bayesian assignment analysis revealed two genetic groups, while the genetic structure of the populations could not be explained by isolation‐by‐distance. Genetic and fatty acid profile data were not correlated based on Mantel test. Also, phorbol ester content and genetic diversity were not associated. Multiple linear regression analysis showed that total oil content was associated with altitude and seasonality of temperature. The content of unsaturated fatty acids was associated with altitude. Therefore, the cultivation planning of J. curcas should take into account chemical variation related to environmental factors.     

麻风树LEC1基因的生物信息学分析

LEC1是调控植物种子发育过程和油脂产量的重要基因。本文选择功能已知的拟南芥LEC1基因为参考序列,对麻风树LEC1基因开展了蛋白组分和理化性质分析,蛋白结构和功能预测,系统进化分析等生物信息学研究,为进一步研究麻风树LEC1基因的生物学功能奠定了基础。     

Biodiesel production from high acid value waste frying oil catalyzed by superacid heteropolyacid

Transesterification of waste cooking oil with high acid value and high water contents using heteropolyacid H3PW12O40 x 6H2O (PW12) as catalyst was investigated. The hexahydrate form of PW(12) was found to be the most promising catalyst which exhibited highest ester yield 87% for transesterification of waste cooking oil and ester yield 97% for esterification of long-chain palmitic acid, respectively. The PW12 acid catalyst shows higher activity under the optimized reaction conditions compared with conventional homogeneous catalyst sulfuric acid, and can easily be separated from the products by distillation of the excess methanol and can be reused more times. The most important feature of this catalyst is that the catalytic activity is not affected by the content of free fatty acids (FFAs) and the content of water in the waste cooking oil and the transesterification can occur at a lower temperature (65 degrees C), a lower methanol oil ratio (70:1) and be finished within a shorter time. The results illustrate that PW12 acid is an excellent water-tolerant and environmentally benign acid catalyst for production of biodiesel from waste cooking oil.     

麻疯树叶中过氧化物酶JCP-1的分离纯化和几种特性的检测

麻疯树叶片蛋白粗提液经硫酸铵分级沉淀,强阴离子琼脂糖、强阳离子琼脂糖和交联葡聚糖层析,得到一个比活为4499U·mg-1（蛋白）过氧化物酶,命名为JCP-1。其分子量为49kDa,等电点为pH3.3,最适pH为5.0-6.0。以H2O2为底物的Km为2.14mmol·L-1。JCP-1具有宽泛的最适保存pH（7.0-11.0）和较高的耐热性（80℃高温处理15min,活性保持在90%以上）。30%PEG6000处理模拟干旱胁迫及50℃高温胁迫麻疯树苗,其叶片中JCP-1活性分别提高121%和155%。     

Isolation of novel microsatellites from Jatropha curcas L. and their cross-species amplification

Jatropha curcas L., a member of the Euphorbiaceae, is widely distributed in different parts of the globe. In the present study, 12 microsatellites were isolated from J. curcas and their cross-species amplification was checked in six species of genus Jatropha. Within J. curcas, observed and expected heterozygosities ranged from 0.94 to 0.54 and from 0.95 to 0.56, respectively. Of the 12 loci, five showed significant deviation from Hardy-Weinberg equilibrium. There was no significant linkage disequilibrium detected between any of the loci. The markers isolated in the present investigation will be useful for assessing the population diversity and genetic structure of J. curcas and also in other species of Jatropha.     

Mechanistic modeling of biodiesel production using a liquid lipase formulation

In this article, a kinetic model for the enzymatic transesterification of rapeseed oil with methanol using Callera? Trans L (a liquid formulation of a modified Thermomyces lanuginosus lipase) was developed from first principles. We base the model formulation on a Ping‐Pong Bi‐Bi mechanism. Methanol inhibition, along with the interfacial and bulk concentrations of the enzyme was also modeled. The model was developed to describe the effect of different oil compositions, as well as different water, enzyme, and methanol concentrations, which are relevant conditions needed for process evaluation, with respect to the industrial production of biodiesel. The developed kinetic model, coupled with a mass balance of the system, was fitted to and validated on experimental results for the fed‐batch transesterification of rapeseed oil. The confidence intervals of the parameter estimates, along with the identifiability of the model parameters were presented. The predictive capability of the model was tested for a case using 0.5% (wt. Enzyme/wt. Oil), 0.5% (wt. Water /wt. Oil) and feeding 1.5 times the stoichiometric amount of methanol in total over 24 h. For this case, an optimized methanol feeding profile that constrains the amount of methanol in the reactor was computed and the predictions experimentally validated. Monte‐Carlo simulations were then used to characterize the effect of the parameter uncertainty on the model outputs, giving a biodiesel yield, based on the mass of oil, of 90.8 ± 0.55 mass %. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:1277–1290, 2014     

Chemical Constituents from Roots of Jatropha curcas

Thirteen compounds were isolated from the roots of datropha curcas L. Combining the determination of physico-chemical constants and spectral analyses (IR, 1H-NMR, 13C-NMR, EIMS, FABMS), the structures of the compounds were identified as 5α-stigmastane-3, 6-dione (1), nobiletin (2), β-sitosterol (3), taraxerol (4), 2S-tetracosanoic acid glyceride-1(5),5-hydroxy-6,7-dimethoxycoumarin (6), jatropholone A (7), jatropholone B (8), 6-methoxy-7-hydroxycoumarin (9), caniojane (10), 3-hydroxy-4-methoxybenzaldehyde (11), 3-methoxy-4-hydroxybenzoic acid (12) and daucosterol (13). Among them, compound 5 is a new compound which has never been reported in China and abroad, compound 1, 2, 9, 10, 11, 12 were first time isolated from the plant, 7 and 8 are a pair of stereoisomers which can be inverted in dilute basic solution. 10 is a diterpenoid containing peroxide bridge.