The Kinetics of Interesterfication on Waste Cooking Oil (Sunflower Oil) for the Production of Fatty Acid Alkyl Esters using a Whole Cell Biocatalyst (Rhizopus oryzae) and Pure Lipase Enzyme

Recent research and technology have provided promising outcomes to rely on biodiesel as the alternative and conventional source of fuel. The use of renewable sources constitutes the main stream of research. Waste Cooking Oil (WCO) was used for biodiesel production in this study. Lipase enzyme producing fungi Rhizopus oryzae 262 and commercially available pure lipase enzyme were used for comparative study in the production of FAAE. The whole cell biocatalyst and pure enzyme were immobilized using calcium alginate beads. It was prepared by optimizing with different molar ratios of calcium chloride and different percent sodium alginate. Entrapment immobilization was done for whole cell biocatalyst. PE was also immobilized by entrapment for the transesterification reaction. Four different solvents methanol, ethanol, n-propanol, n-butanol were used as the acyl acceptors. The reaction parameters like temperature, molar ratio, reaction time, and amount of enzyme to be used were also optimized for methanol alone. The same parameters were adopted for the other acyl acceptors too. Among the different acyl acceptors, methanol whose reaction parameters were optimized showed maximum conversion of triglycerides to FAAE - 94% with PE and 84% with WCB. On the whole, PE showed better catalytic converting ability with all the acyl acceptor compared to WCB, further study, it was observed that three consecutive and reversible reactions occurred in the interesterification of triglycerides. So, a kinetic model based on Michaelis-Menten equation with competitive substrate inhibition was used to find the maximum reaction rate V_i for the four solvents using pure enzyme and WCB.     

米曲霉产氨基酰化酶条件及部分酶学性质研究

通过单因子和多因子摇瓶正交优化试验,确定了米曲霉液态发酵产氨基酰化酶的最佳发酵条件.优化发酵培养基组成(ρ/gL-1):葡萄糖40,蔗糖10,可溶性淀粉20,蛋白胨2.5,马铃薯液1000mL,pH自然.培养基装量50mL/250mL三角瓶,接种量4%.培养温度30℃,转速100r/min,发酵时间42h.每50mL培养物的总酶活由优化前的2627u提高到7338u,是优化前的2.79倍.研究了米曲霉氨基酰化酶的部分酶学性质.该酶催化反应的最适pH为7.0,最适温度为40℃,低浓度的Co2 (5×10-4mol/L)对酶活激活作用显著.图5表2参8     

黄单胞菌生物滴滤塔处理硫化氢能力及产物研究

以黄单胞菌为硫氧化菌种挂膜于生物滴滤塔,研究其去除能力及产物.通过测定压降、生物量确定挂膜时间,以进出气口浓度变化反映去除效率,测定营养液pH值变化,用SEM电镜扫描、能谱分析、X射线衍射对产物进行分析.结果表明,黄单胞菌挂膜时间为4 d,持续运行,菌种均匀地分布于填料表面;保持气体流量为1 L/min,H_2S气体质量浓度在2 000 mg/m~3以下时,去除率大于90.5%,单位体积最大生化去除量为43.23 g/(m~3·h);以10 d为周期添加营养,系统的pH值在5～7;自然沉降得到的黄色颗粒状沉淀粒径为3.161～31.90 μm;能谱分析表面主要成分为C、O、S,应该为微生物和S系物;X射线衍射分析表明干燥粉末的主要无机物质存在形态为S单质.塔在弱酸环境运行,能减少对装置的腐蚀,营养液中通过物理沉降、过滤、低速离心的简单手段,就能回收产物S单质.     

Controlling bacterial leaf blight of rice and enhancing the plant growth with endophytic and rhizobacterial Bacillus strains

This study was conducted to assess efficacy of biological control against bacterial leaf blight (BLB) of rice produced by Xanthomonas oryzae pv. oryzae. Five endophytic strains (A1, A2, A3, A13 and A15) and two rhizospherial Bacilli (D29 and H8) were tested for their antagonistic activities against BLB in vitro and in vivo. All seven strains showed high potential of antagonistic activity against X. oryzae pv. oryzae and three phytopathogenic fungi in vitro. Test of 16SrRNA gene sequence were assigned isolates A1, A3 and A13 as Bacillus amyloliquefaciens while isolates A2 and A15 as B. methylotrophicus and B. subtilis, respectively. In greenhouse, four strains of displayed 50.29%–57.86% inhibition rate against the pathogen and significantly increased plant fresh weight from 50.03% to 73.11% and dry weight from 64.11% to 86.65% in treated rice plants. In addition, these strains demonstrated strong capability to produce indole-3-acetic acid, siderophores, solubilizing phosphate and also colonize roots. Real-time quantitative polymerase chain reaction revealed that expression of defense-related genes including OsAOS2, OsJMT1, OsNPR1 and OsPR1b were significantly up-regulated in leaves of D29-exposed rice plants, suggesting that treatment of rice with D29 suppressed BLB through systemic activation of the plant defense system. Therefore, data suggest that Bacillus isolates A13, A15, D29 and H8 support effective antagonistic activity against BLB under greenhouse conditions in addition to their potential to promote growth of rice plants.     

两级滴滤去除硫化氢和甲硫醇混合恶臭气体

把氧化硫硫杆菌(T. thiooxidans)、排硫硫杆菌(T. thioparus)组成的自养菌群和黄单胞菌(Xanthomonas)为主的异养菌群分别接种在两个生物滴滤反应器中,将其依次串联净化处理硫化氢(H₂S)、甲硫醇(MT)混合臭气.一级反应器(A^#)的复合自养菌在酸性环境下对负荷为7～8g/(m³h)的H₂S平均去除率可达94%,且不受混合气体中MT含量的影响.二级反应器(B^#)适于中性环境,对负荷为4～5g/(m^{3h)的MT平均去除率为83%.若H₂S在混合气体中浓度过高,经A#处理后的浓度仍高于50mg/m3,可导致后反应器酸化,使MT脱臭效率显著下降.}     

Inhibition mechanism of fermentation broth extract of Phomopsis sp. Strain S4 on cell membranes of Magnaporthe oryzae北大核心CSCD

The secondary metabolites of endophytic Phomopsis sp. strain S4 show antifungal activity against a variety of plant pathogens, which implies that strain S4 has potential prospect in crop disease control. The aim of this study was to explore the inhibition mechanisms of S4 against plant pathogenic fungi. Magnaporthe oryzae was used as the main pathogenic material for the research. Cell membrane changes were detected using antifungal experiments, scanning electron microscopy, Q RT-PCR, and cell content leaking experiments. The results of scanning electron microscopy showed that M. oryzae mycelia, after treatment with S4 fermented product extract, decreased in size, suggesting the integrity of the cell membrane was destroyed. The genes related to ergosterol synthesis, which plays an important role in membrane integrity, were studied though Q RT-PCR. The results showed that the expression levels of ERG1, ERG11, and ERG6 genes were down-regulated by 2.0, 1.40, and 2.7-fold, respectively, whereas that of ERG7 was up-regulated by 1.38-fold, which means the ergosterol synthesis pathway was destroyed. The physiological experiments also showed that the cell contents of M. oryzae mycelia treated with S4 fermented product extract leaked significantly, which was consistent with the Q RT-PCR results. The results showed that S4 fermentation broth exact could destroy the cell membrane integrity by inhibiting ergosterol synthesis, and eventually inhibit M. oryzae cell growth. © 2018 Science Press. All rights reserved.     

Synthesis of 2-furyl-4-arylidene-5(4H)-oxazolones as new potent antibacterial agents against phyto-pathogenic and nitrifying bacteria

Crop losses due to bacterial pathogens are a major global concern. Most of the available pesticides for these pathogens suffer from various drawbacks such as complicated synthesis, high cost, high toxicity, pesticide resistance and environmental hazards. To overcome these drawbacks, the present study was undertaken to find a potent bactericide. Therefore, a series of compounds comprising bioactive furyl and oxazolone rings was synthesized under microwave irradiation and screened for in vitro antibacterial activity. The reactions were completed in fewer than 2 minutes with minimal use of solvents and resulted in high yields. These compounds were screened for antibacterial activity against plant pathogens, Xanthomonas oryzae, Ralstonia solanacearum and nitrifying bacteria, Nitrosomonas species under laboratory conditions. Five compounds were active as antibacterial agents against Xanthomonas oryzae and Ralstonia solanacearum. However, all compounds were effective against the Nitrosomonas species and the best one was 2-furyl-4-(3-methoxy-4-hydroxybenzylidene)-5(4H)-oxazolone. The study revealed the fast and environmentally friendly synthesis of bioactive title compounds, which also hold promise to be used as prototypes for the discovery of potent analogues.     

抗柑桔溃疡病菌可溶性单链抗体的表达及鉴定

前期采用核糖体展示技术筛选了抗柑桔溃疡病菌(Xanthomonas axonopodis pv.citri,Xac)高亲和力单链抗体(Single chain variable fragment,scFv)GX13、GX44和GX95,为高效表达具有生物活性的单链抗体,本研究将前期筛选的高亲和力单链抗体基因转入大肠杆菌HB2151中进行可溶性表达,点印迹和Western印迹检测可溶性单链抗体的表达水平,亲和层析纯化表达的单链抗体,酶联免疫吸附法(ELISA)检测纯化单链抗体的抗原结合活性.结果显示,3株抗柑桔溃疡病菌可溶性单链抗体在大肠杆菌HB2151中均获得成功表达,表达产物分子量(Mr)约为32.0×103,主要集中于细菌周质腔中.ELISA检测纯化的单链抗体都具备抗原结合活性,可进一步应用于柑桔溃疡病菌的免疫诊断和病害防治.图4参14     

米曲霉氨基酰化酶酶法拆分制备D型芳香族氨基酸

以DL-苯丙氨酸为起始原料,乙酸酐乙酰化后得到N-乙酰DL-苯丙氨酸,利用米曲霉氨基酰化酶立体结构专一催化水解N-酰基-L-氨基酸的酰胺键的特点,分别得到L-苯丙氨酸和N-乙酰D-苯丙氨酸.通过结晶法分离两者后,N-乙酰D-苯丙氨酸经6 mol/L HCI水解得到D-苯丙氨酸.由此拆分DL-苯丙氨酸得到光学纯度分别为98%的L-苯丙氨酸(收率84.8%,以原料N-乙酰-DL-苯丙氨酸的0.5倍摩尔量为理论产率)和92.3%的D-苯丙氨酸(收率89.5%).参22