枯草芽孢杆菌感受态细胞的制备及质粒转化方法研究

为便于枯草芽孢杆菌工业化生产应用,对Spizizen创立的枯草芽孢杆菌DNA转化方法进行改进.用GMI和GMII溶液处理枯草芽孢杆菌野生型菌株BS501a、营养缺陷型突变株DBl342和非营养缺陷型突变株WB800,用改进的方法制备感受态细胞,用7.5kb质粒pSBPTQ进行转化,并研究RNA、酵母粉、水解酪蛋白、培养方法对枯草芽孢杆菌质粒转化的影响.结果表明,该方法适用于不同基因型枯草芽孢杆菌的质粒转化,营养缺陷型突变株DBl342的转化率为750 CFU/μg/DNA,非营养缺陷型突变株WB800转化率为1 070 CFU/xg DNA,野生型菌株BS501a转化率为270 CFU/μg/DNA.根据影响转化效率的因素,推测在该方法中,枯草芽孢杆菌质粒转化原理:一定生物量的枯草芽孢杆菌在外界营养条件和钙、镁离子作用下,细胞壁和细胞膜形成缺陷,使外源DNA转入枯草芽孢杆菌细胞内.     

肌苷生产菌枯草芽孢杆菌ATCC 13952的全基因组测序及序列分析

摘要:【目的】枯草芽孢杆菌ATCC 13952是一株肌苷工业生产菌株。为深入研究ATCC 13952菌株积累肌苷的分子机制以及为进一步分子育种研究提供序列背景信息,有必要解析ATCC 13952菌株的基因组序列信息。【方法】本研究采用高通量测序和Sanger测序相结合对ATCC 13952菌株进行全基因组测序,然后使用相关软件对测序数据进行基因组组装、基因预测与功能注释、GO/COG 聚类分析、共线性分析等。【结果】枯草芽孢杆菌ATCC 13952整个基因组大小为3876276 bp,GC含量为45.8%,序列已提交至GenBank 数据库,登录号为CP009748。比较基因组及嘌呤代谢相关基因分析结果显示:枯草芽孢杆菌ATCC 13952与其他几株芽孢杆菌具有较好的基因组共线性关系,嘌呤代谢相关基因编码的蛋白与标准菌株比较发生了一些缺失和突变。【结论】本研究首次报道了一株肌苷生产菌枯草芽孢杆菌ATCC 13952的全基因组序列,分析了基因组基本特征,初步探讨了该菌株积累肌苷的分子机制,为后续的进一步分子育种提供了理论基础。     

3-甾酮-1-脱氢酶基因的表达及甾体转化分析

本文利用带有P43启动子的表达分泌载体pWB980,实现了简单节杆菌3-甾酮-1-脱氢酶在枯草芽孢杆菌中的表达,表达出的目的蛋白的分子量为55KDa。利用分光光度法检测得到胞内和胞外可溶性部分的酶活分别为110±0.5mU和15±0.6mU每毫克蛋白, 比出发菌株简单节杆菌提高了将近30倍。重组芽孢杆菌对甾体底物4-AD的转化率为45.3％,比出发菌株简单节杆菌提高了近10倍。利用枯草芽孢杆菌对甾体底物进行脱氢为甾体药物的生产开辟了一个新的途径。     

Treatment of poplar alkaline peroxide mechanical pulping (APMP) effluent with Aspergillus niger

Although the moderate load (COD of 5000-10,000 mg/L) and biodegradability of the APMP pulping effluent should allow biological treatment, toxic compounds in the effluent can interfere with this type of treatment. Studies were conducted to determine if treatment of the effluent with Aspergillus niger S13 was feasible. Under the optimized conditions (3% inoculum, pH 6, shaking at 160 rpm, 60-72 h, and 30 °C), this fungus was able to remove about 97% of the methyl tertiary butyl ether (MTBE) extractives, and 60%, 77% and 43% of the chemical oxygen demand, turbidity and color even without a pre-flocculation step. These results are of practical interest in China because the APMP process has become popular, and efficient and cost-effective effluent treatment technologies are in high demand.     

棉花秸秆催化热解特性及动力学的研究

通过热重分析实验观察K2CO3、KOH、KCl、NaCl、MgCl2和ZnCl26种无机催化剂对棉花秸秆热解催化效果的影响。K2CO,、KOH处理过的棉花秆与纯棉花秆相比热解发生在较低的温度范围,而KCl、NaCl、MgCl,和ZnCl2处理过的棉花秆热解发生在较高的温度范围。碱性催化剂K2C03、KOH降低了棉花秸秆的最大质量损失率,而KCl、NaCl、MgCl2和ZnCl2却增大了棉花秸秆的最大质量损失率。应用Ozawa动力学模型得到动力学参数,棉花秸秆在热解主要阶段可由一段一级反应过程描述,升温速率10K／min时活化能值EA的范围是35～66kJ／mol,频率因子自然对数的范围是4～12。     

Effect of hot-water extraction on alkaline pulping of bagasse

The effect of hot-water extraction on alkaline pulping was investigated. The properties of black liquor and pulp strength of bagasse were analyzed. The extraction was conducted at 160 °C for 30 min where 13.2% of the mass was dissolved in the extraction liquor. Untreated bagasse and extracted bagasse were digested by soda and soda-AQ processes at 17% and 15.5% (with 0.1% AQ) alkali charge (NaOH). Cooking temperatures were 160 °C and 155 °C respectively. The pulp from extracted bagasse had a lower Kappa number and a higher viscosity compared to the pulp from the untreated bagasse. The black liquor from pulping extracted bagasse had a lower solid content, a lower viscosity and a lower silica content, but a higher heating value than that from pulping of untreated bagasse. Hot-water extraction resulted in a significant decrease in bleaching chemical consumption and the formation of chlorinated organics. Pulp strength properties such as the tensile index and the burst index were found to be lower, but the tear index, bulk, opacity and pulp freeness were found to be higher when hot-water extraction was applied.     

Evaluation of cotton stalk hydrolysate for xylitol production

Cotton stalk is a widely distributed and abundant lignocellulosic waste found in Turkey. Because of its rich xylose content, it can be a promising source for the production of xylitol. Xylitol can be produced by chemical or biotechnological methods. Because the biotechnological method is a simple process with great substrate specificity and low energy requirements, it is more of an economic alternative for the xylitol production. This study aimed to use cotton stalk for the production of xylitol with Candida tropicalis Kuen 1022. For this purpose, the combined effects of different oxygen concentration, inoculum level and substrate concentration were investigated to obtain high xylitol yield and volumetric xylitol production rate. Candida tropicalis Kuen 1022 afforded different concentrations of xylitol depending on xylose concentration, inoculum level, and oxygen concentration. The optimum xylose, yeast concentration, and airflow rate for cotton stalk hydrolysate were found as 10.41 g L^?1, 0.99 g L^?1, and 1.02 vvm, respectively, and under these conditions, xylitol yield and volumetric xylitol production rate were obtained as 36% and 0.06 g L^?1 hr^?1, respectively. The results of this study show that cotton stalk can serve as a potential renewable source for the production of xylitol.     

Impact of fly ash pretreatment on aerobic treatment of thermomechanical pulping spent liquor

This study investigated the impact of biomass‐based fly ash (FA) pretreatment on the biodegradability of a thermomechanical pulping spent liquor (TMPL) in an aerobic system. In this study, FA was mixed with TMPL under the conditions of 6 wt.% based on TMPL, 25°C and 10 h, which removed a part of recalcitrant organic materials and resulted in 68.0, 40.0, 60.1, 81.2 and 48.3% reductions in chemical oxygen demand (COD), biochemical oxygen demand (BOD), total organic carbon (TOC), lignin and sugar, respectively. FA‐pretreated TMPL pressate (FA‐TMPL) was biologically treated in an aerobic system of sequencing batch reactor (SBR). The performance of the biological treatment with and without FA pretreatment was studied in two parallel SBRs over three months. The combination of FA and biological treatments removed 97.3% of COD, 98.3% of BOD, 96.3% of lignin, 99.5% of sugar, and 98.1% of TOC. Without FA pretreatment, the biological system removed 87.3% of COD, 89% of BOD, 81.6% of lignin, 98.6% of sugars, and 90.5% of TOC. The results also confirmed that the settling ability of sludge, which was indicated as a sludge volume index, was reduced from 109.3 mL/g to 53.5 mL/g. In addition, the advantages of using FA pretreatment in aerobic systems were discussed in detail. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:370–378, 2018     

Biodegradation of exploded cotton stalk by Bacillus sp.

The exploded bast, branch and stem of cotton stalk were degraded by alkalophilic Bacillus NT-19, with weight losses of 24%, 20% and 14%, respectively, after 14 d. Compared with a white-rot fungus (Phanerochaete chrysosporium), Bacillus NT-19 preferentially degraded the non-cellulose components of cotton stem. The relative degree of crystallinity of bast fibers decreased by 8% and the middle lamella was partially removed from the fiber bundle by the Bacillus.     

Characterisation of the heterogeneous alkaline pulping kinetics of hemp woody core

A model was developed, based on the power law of growth and Avrami's concepts in nuclei growth to describe the heterogeneous nature of alkaline pulping kinetics, taking into account the effects of effective alkali concentration and temperature. It was then applied against published data to estimate model parameters. The final form of the model applied to alkaline pulping of thin hemp woody core in flow-through reactors could be represented by a first order rate equation with a time-dependent rate constant: [-dY/dt = A.[OH-]b.exp (-E/RT)n.Y.t(n-1). The rate equations applied for delignification, cellulose and xylan losses. It was found that the heterogeneous kinetic models predicted well the delignification and xylan loss of hemp woody core, with R2 values of 0.96 and 0.97, respectively. The model was, however, less accurate in predicting cellulose loss. The values n, which could represent the acceleration/deceleration extent of growth of reacting volumes, were found to be less than unity. This demonstrated the heterogeneous characteristics of alkaline pulping for hemp woody core.     

Biotransformation of banana waste into protein by pleurotus sajor-caju

Pseudostems and leaves from banana waste were used for biotransformation into protein by using P. sajor-caju, an oyster mushroom. Treatment of formalin (500 ppm) + carbendazim (12.5 ppm) of these substrates was found to favour relatively high percentage biological efficiency (BE) of P. sajor-caju.. Steam sterilization also exhibited comparable yield performance by P. sajor-caju. Fruiting bodies harvested from all the treatments had relatively higher protein contents. The spent substrate (steam sterilized) was found to be suitable as an ideal animal feed because of its rich nutritive composition.     

秸秆还田对棉花生长的化感效应

研究了秸秆不同还田量及不同腐解时间对后茬棉花抗氧化物酶活性和光合生理特性的影响.结果表明,棉花秸秆腐解产物,在一定程度上抑制了棉花种子的萌发,延长了种子出苗时间,影响棉花植株生长.随着秸秆腐解时间延长和秸秆还田数量增加,棉花单叶净光合速率、气孔导度、蒸腾速率和胞间CO2浓度降低.当加入秸秆量为10 g·kg-1时,净光合速率略高于对照,随着秸秆还田量的增加,又呈现下降趋势,表现出一定"浓度效应",其中加入秸秆量为90 g·kg-1时,净光合速率下降最明显,腐解30d和60d净光合速率分别比对照减少了26.8%和43.4%;随着秸秆还田量的增多和秸秆腐解时间延长,POD活性增加,SOD活性和根系活力呈现下降趋势,叶片MDA含量却升高.表明棉花秸秆还田后分解产生的化学物质具有一定的自毒效应,对连作棉花种子萌发、抗氧化物酶活性和光合生理造成影响.