葡萄糖及磷酸盐对刺糖多孢菌多杀菌素发酵的影响

The effects of glucose and inorganic phosphate on mycelium growth and spinosad production with Saccharopolyspora spinosa were studied. The results showed that the increase of glucose concentration from 18.6g·L-1 to 58.8g·L-1 could promote the mycelium growth and spinosad production. And when the glucose concentration increased from 58.8g·L-1 to 79.6g·L-1, no obvious change was detected but a slight drop in spinosad production was observed, whereas, when the glucose concentration increased from 79.6g·L-1 to l15.3g·L-1, substantial decrease in both mycelium growth and spinosad production occurred. The increase of phosphate conccntration from 3.68mmol· L-1 to 29.41mmol·L-1 rendered corresponding increase in mycelium growth and spinosad production. When phosphate concentration increased from 29.41mmol· L-1 to 44.12mmol·L-1, mycelium growth slightly increased and spinosad production dropped, while when phosphate concentration increased from44.12mmol·L-1 to 57.62mmol·L-1, both mycelium growth and spinosad production decreased sharply. Conclusively,the optimal initial concentration of glucose and phosphate is 58.8g·L-1 and 29.41mmol·L-1, respectively. The spinosad fermentation in the production medium containing 58.8g·L-1 glucose and 29.41mmol·L-1 phosphate was scaled up in 5-L fermentation and the spinosad production reached 507mg·L-1, which was 28% higher than that in the flask fermentation.     

酒精-沼气双发酵耦联工艺中丙酸对酿酒酵母酒精发酵的影响(英文)

Propanoic acid accumulated in an ethanol-methane coupled fermentation process affects the ethanol fermentation by Saccharomyces cerevisiae. The effects of propanoic acid on ethanol production were examined in cassava mash under different pH conditions. Final ethanol concentrations increased when undissociated propanoic acid was <30.0 mmol·L-1 . Propanoic acid, however, stimulated ethanol production, as much as 7.6% under proper conditions, but ethanol fermentation was completely inhibited when undissociated acid was >53.2 mmol·L-1 . Therefore, the potential inhibitory effect of propanoic acid on ethanol fermentation may be avoided by controlling the undissociated acid concentrations through elevated medium pH. Biomass and glycerol production decreased with propanoic acid in the medium, partly contributing to increased ethanol concentration.     

ETHANOL FERMENTATION OF MIXED GLUCOSE AND XYLOSE BY PICHIA STIPITIS

Pichia stipitis CBS 5773 yeast cells were used to ferment the mixed substrates consisted of glucose andxylose to produce ethanol.The effects of aeration rate,initial substrate concentration and pH on substrateutilization and ethanol yield were evaluated.During batch fermentation,the oscillation phenomena in cell growthwere observed at low aeration rate,whereas the diauxic growth at high aeration rate.The substrate utilizationratio and ethanol yield reached 95％ and 0.46g/g respectively under appropriate operation conditions.Amodified unstructural model was proposed to simulate the diauxic cell growth,substrate consumption andproduct formation.     

Rhodococcus sp. SHZ-1腈水合酶的诱导表达和稳定性研究

Inducing expression and the reaction characteristic of nitrile hydratase (NHase) from Rhodococcus sp.SHZ-1 were investigated. The results showed that the expression of NHase was greatly enhanced with the cooperation of acrylonitrile and ammonium chloride as inducer in the medium and the specific activity of NHase was increased of 44%. Then the temperature, pH, concentration of acrylonitrile and acrylamide were evaluated, which affected the activity and reaction characteristic of NHase. It was found that the temperature and concentration of acrylamide were the most important factors for the catalyzation of NHase. The optimal catalysis temperature of NHase from Rhodococcus sp. SHZ-1 was 30℃, and the activation energy of the hydration of NHase was 90.2kJ·mol-1 in the temperature range from 5℃ to 30℃. Km of NHase was 0.095mol.L-1 using acrylonitrile(AN)as substrate, and NHase activity was inhibited seriously when acrylonitrile concentration was up to 40g·L-1, the substrate inhibition constant Ki is 0.283mol·L-1. Moreover, the NHase from Rhodococcus sp. SHZ-1 had very strong tolerance to acrylamide, in which the final concentration of acrylamide reached to 642g·L-1 and the residual activity of NHase still maintained 8.6% of the initial enzyme activity.     

渗透压调节剂对耐高渗菌Candida Krusei生产甘油的影响

Candida krusei was osmotolerant yeast for the production of glycerol. Extracellular osmotic pressure was one of the key factors to induce the enzyme activity of glycerol-3-phosphate dehydrogenase（GPDH） which severely affected the glycerol productivity. Three osmoregulators such as NaCl, PEG4000 and glycerol were used to investigate their effects on yeast growth, glucose consumption and glycerol production. In order to determine the effect of extracellular glycerol concentration, different amounts of glycerol were initially supplemented as an osmoregulator to increase glycerol production. The maximum glycerol concentration attained 179g·L^-1 with initial glycerol concentration of 80g·L^-1 in medium, compared with 41g·L^-1 in the control experiment. These results were successfully reoroduced for fed-batch orocess in an air-lift reactor.     

利用农业废弃物和诱导剂提高海洋内生拟盘多毛孢菌J63的漆酶产量(英文)

Pestalotiopsis sp. J63, producing a high activity of laccase, is a new marine-derived fungus isolated from the oceanic sediment of the East China Sea. Since the marine environment is oligotrophic nutrient, marine de-rived fungi may use small amount of nutrients to grow and produce laccases. Agricultural residues that are mainly composed of lignin, cellulose and hemicellulose are difficult to be degraded and few microbes can take them as sub-strates, so they are considered as oligotrophic nutrient and have the potential to be used to produce value added products. In this study, the ability of Pestalotiopsis sp. J63 to use agricultural residues to produce laccases was tested in the submerged fermentation. The combination of 3 g·L-1 maltose and 20 g·L-1 rice straw was the best car-bon sources and 8 g·L-1 ammonium sulfate was the best nitrogen source under the condition without inducers. The effects of five inducers, the feeding time and concentration of inducer on laccase production were investigated. Adding 0.09 mmol·L-1 phenol after 24 h of incubation led to high laccase activity （5089 U·L-1）, while with 0.09 mmol·L-1 phenol in the medium and wheat bran as the nitrogen source, the laccase activity could reach 5791.7 U·L-1. Native-PAGE results showed that two laccase isozymes were present in the cultures. One existed in both in-duced and non-induced culture filtrates, while the other was only found in the fermentation with the addition of phenol, guaiacol and veratryl alcohol.     

A novel aeration strategy in repeated-batch fermentation for efficient ethanol production from sweet sorghum juice

To improve the efficiency of ethanol production in a batch fermentation from sweet sorghum juice under a very high gravity(VHG) condition(～ 290 g/L of total sugar) by Saccharomyces cerevisiae NP01, repeatedbatch fermentation under an aerated condition(2.5 vvm for the first 4 h during every cycle) was done in a5-L fermenter. The average ethanol concentration(P), productivity(Qp) and yield(Yp/s) for five successive cycles were 112.31 g/L, 1.55 g/L·h~(-1) and 0.44, respectively with 80.97% sugar consumption. To complete sugar consumption, the total sugar of the juice was reduced to a high gravity(HG) level(～240 g/L). The results showed that yeast extract was not necessary for ethanol production, and aeration during every other cycle i.e., alternating cycles, was sufficient to promote both yeast growth and ethanol production.The average P, Qpand Yp/svalues for eight successive cycles with aeration during alternating cycles were97.58 g/L, 1.98 g/Láh and 0.41, respectively with 91.21% sugar consumption. The total fatty acids in the yeast cells under the aerated condition were ～ 50% higher than without aeration, irrespective the initial sugar concentration, whereas the ergosterol contents under aeration condition were ～ 29% to 49% higher than those without aeration.     

Optimization of hydrothermal pretreatment for co-utilization of xylose and glucose of cassava anaerobic residue for producing ethanol

The development of a process that could recover biofuel from industrial cellulose waste can not only reduce the negative environmental impacts by using fossil fuels, but also bring a green idea for the waste's disposing. In this study, hydrothermal pretreatment was optimized for cassava anaerobic residue, a cellulosic waste from cassava ethanol industry, to co-utilize xylose and glucose for producing bioethanol. The effect of the main pretreatment conditions, namely, temperature, solid content and time, was explored for the highest recovery of xylose in prehydrolysate and glucose in enzymatic hydrolysate. The single factor experiment results showed that the conditions for maximum xylose recovery in prehydrolysate and glucose recovery in enzymatic hydrolysate were 60 °C, 75 min, 10% solids and 160 °C, 75 min, 10% solids, respectively. Whereafter, response surface methodology(RSM) was applied to further optimize the pretreatment conditions for the maximum theoretical ethanol production through utilizing both xylose and glucose. A treatment at 163 °C, for 59 min and with 9.5%solids was found optimal, with the highest ethanol production of 20.2 mg·g~(-1) raw material. Furthermore, in order to assess the impacts of the pretreatment on cassava anaerobic residue, the changes in crystallinity and morphology for untreated and pretreated solids were investigated.     

运用试验读者设计方法提高胞外杂合β-葡聚糖酶在重组大肠杆菌中的表达

A genetically engineered Escherichia coli JM109 harboring pLF3 was used to produce a hybrid extracellular β-glucanase. Starting with enzyme production medium, glycerol and yeast extract combined with NaNO3 were screened to be the most suitable carbon and nitrogen source, respectively. Analysis of six components of the enzyme production medium by employing statistical optimization methods such as Plackett-Burman design and steepest ascent showed that yeast extract was the only significant variable and its best concentration for enzyme production was 12g·L-1. After optimization of the medium, 297.71U·ml-1 of β-glucanase activity in the medium and 352350U·g-1 of β-glucanase selectivity could be obtained, which were 14 and 72 folds higher than those obtained from original medium, respectively. Even higher enzyme activities were achieved by batch cultivations in a conventional stirred bioreactor on the optimized medium.     

Kinetics of glucose ethanolysis catalyzed by extremely low sulfuric acid in ethanol medium简

The kinetics for production of ethyl levulinate from glucose in ethanol medium was investigated. The experiments were performed in various temperatures （433-473 K） and initial glucose concentrations （0.056-0.168 mol·L-1） with extremely low sulfuric acid as the catalyst. The results show that higher temperature can improve the conversion of glucose to ethyl levulinate, with higher yield of ethyl levulinate （44.79%, by mole） obtained at 473 K for 210 min. The kinetics follows a simplified first-order kinetic model. For the main and side reactions, the values of activation energy are 122.64 and 70.97 kJ·mo1-1, and the reaction orders are 0.985 and 0.998, respectively.     

气提法弱化超高浓度乙醇发酵过程中自絮凝酵母振荡行为

超高浓度(very high gravity,VHG)连续乙醇发酵过程中的振荡行为会导致发酵终点的乙醇浓度振荡和降低,是VHG连续乙醇发酵面临的主要问题。研究表明,游离酵母Saccharomyces cerevisiae 4126和自絮凝酵母BHL01在VHG连续乙醇发酵过程中,生物量、残糖、乙醇、甘油等发酵参数都呈现130～145 h的周期性振荡,且絮凝酵母发酵体系的平均乙醇浓度和乙醇生产强度都明显高于游离酵母体系。在絮凝酵母VHG连续乙醇发酵过程中利用发酵尾气气提分离乙醇,酵母细胞振荡行为被明显弱化,达到拟稳态状态,并且使发酵液中的残糖浓度控制在0.1 g·L^-1以下,平均乙醇浓度为110.87 g·L^-1,乙醇产率达到2.99 g·L^-1·h^-1。因此,本研究为弱化VHG连续乙醇发酵中的参数振荡行为提供了新的技术手段。     

氧化还原电位控制的自絮凝酵母自动重复批次发酵

引言随着石油资源的日益减少和环境污染的逐渐加剧,使用可再生的清洁能源已经是世界各国的共识。在众多形式的非矿物质能源中,基于生物质的燃料乙醇已得到了广泛的应用[1]。但是,由于生产成本较高,各国的燃料乙醇生产大都依靠着政策扶植和税收优惠[2]。因此,看似十分"成熟"的乙醇生产产业,仍然需要进一步开发降低成本的创新技术[3]。近些年来,通过基因工程手段改造菌种[4]、     

Study of simultaneous saccharification and fermentation for steam exploded wheat straw to ethanol

Although simultaneous saccharification and fermentation (SSF) has been investigated extensively, the optimum condition for SSF of wheat straw has not yet been determined. Dilute sulfuric acid impregnated and steam explosion pretreated wheat straw was used as a substrate for the production of ethanol by SSF through orthogonal experiment design in this study. Cellulase mixture (Celluclast 1.5 l and ?-glucosidase Novozym 188) were adopted in combination with the yeast Saccharomyces cerevisiae AS2.1. The effects of reaction temperature, substrate concentration, initial fermentation liquid pH value and enzyme loading were evaluated and the SSF conditions were optimized. The ranking, from high to low, of influential extent of the SSF affecting factors to ethanol concentration and yield was substrate concentration, enzyme loading, initial fermentation liquid pH value and reaction temperature, respectively. The optimal SSF conditions were: reaction temperature, 35°C; substrate concentration, 100 g·L^-1; initial fermentation liquid pH, 5.0; enzyme loading, 30 FPU·g^-1. Under these conditions, the ethanol concentration increased with reaction time, and after 72 h, ethanol was obtained in 65.8% yield with a concentration of 22.7 g·L^-1.     

过表达谷氧还蛋白基因GRX5提高酿酒酵母乙酸耐性

利用可再生的纤维素原料生产燃料乙醇是国内外研究的热点。但纤维素原料一些预处理过程产生的乙酸对酿酒酵母细胞生长和乙醇发酵产生强烈抑制,因此,提高酿酒酵母细胞的乙酸耐受性是提高纤维素乙醇发酵效率的重要手段。本文研究了谷氧还蛋白家族中GRX5p的编码基因的过表达对酿酒酵母在乙酸胁迫条件下细胞生长和发酵性能的影响。结果表明,过表达GRX5的重组菌株在含有5 g·L^-1乙酸的平板中生长优于对照菌株;在含有5 g·L^-1乙酸的培养基中进行乙醇发酵,过表达GRX5的重组菌株可在48 h基本消耗培养基中所有的葡萄糖,发酵周期比对照菌株缩短了12 h。过表达GRX5菌株的乙醇生产强度为0.897 g·L^-1·h^-1,比对照提高了28.5%。代谢物分析结果表明,过表达GRX5的重组菌株可产生更多的保护性物质海藻糖和甘油,有利于增强菌株胁迫耐受性。     

ORP调控对马克斯克鲁维酵母发酵纤维素水解液的影响

采用通气的方式进行氧化还原电位（oxidation-reduction potential,ORP）调控,研究在添加抑制物条件下Kluyveromyces marxianus 1727-5利用葡萄糖、木糖以及两者混合体系的发酵性能,在此基础上考察了ORP调控策略对玉米秸秆水解液发酵的影响。研究结果表明：在调控ORP策略下,多种混合抑制物对酵母生长代谢造成的损害得以有效改善,细胞活性高,木糖、葡萄糖代谢速率加快。葡萄糖与木糖共发酵时,ORP调控至-150 mV时,葡萄糖发酵时间缩短近30%,木糖醇浓度由3 g·L^-1增加到10 g·L^-1。ORP调控策略也同样能有效缓解玉米秸秆水解液中较高浓度的多种抑制物对酵母细胞的胁迫,ORP为-100 mV时,相比于对照组,在保持终点乙醇不变的情况下,葡萄糖的发酵时间缩短了22%;木糖消耗由4.88 g·L^-1增至10.27 g·L^-1,木糖醇得率也由0.20 g·g^-1提高至0.48 g·g^-1。     

Immobilization of trophic anaerobic acetogen for semi-continuous syngas fermentation

The massive consumption of fossil energy forces people to find new sources of energy. Syngas fermentation has become a hot research field as its high potential in renewable energy production and sustainable development. In this study, trophic anaerobic acetogen Morella thermoacetica was successfully immobilized by calcium alginate embedding method. The ability of the immobilized cells on production of acetic acid through syngas fermentation was compared in both airlift and bubble column bioreactors. The bubble column bioreactor was selected as the better type of bioreactor. The production of acetic acid reached 32.3 g·L^-1 in bubble column bioreactor with a space-time yield of 2.13 g·L^-1·d^-1. The immobilized acetogen could be efficiently reused without significant lag period, even if exposed to air for a short time. A semi-continuous syngas fermentation was performed using immobilized cells, with an average space-time acetic acid yield of 3.20 g·L^-1·d^-1. After 30 days of fermentation, no significant decrease of the acetic acid production rate was observed.     

Efficient acetoin production from pyruvate by engineered Halomonas bluephagenesis whole-cell biocatalysis

Acetoin is an important platform chemical, which has a wide range of applications in many industries. Halomonas bluephagenesis, a chassis for next generation of industrial biotechnology, has advantages of fast growth and high tolerance to organic acid salts and alkaline environment. Here, α-acetolactate synthase and α-acetolactate decarboxylase from Bacillus subtilis 168 were co-expressed in H. bluephagenesis to produce acetoin from pyruvate. After reaction condition optimization and further increase of α-acetolactate decarboxylase expression, acetoin production and yield were significantly enhanced to 223.4 mmol·L^–1 and 0.491 mol·mol^–1 from 125.4 mmol·L^–1 and 0.333 mol·mol^–1, respectively. Finally, the highest titer of 974.3 mmol·L^–1 (85.84 g·L^–1) of acetoin was accumulated from 2143.4 mmol·L^–1 (188.6 g·L^–1) of pyruvic acid within 8 h in fed-batch bioconversion under optimal reaction conditions. Moreover, the reusability of the cell catalysis was also tested, and the result illustrated that the whole-cell catalysis obtained 433.3, 440.2, 379.0, 442.8 and 339.4 mmol·L^–1 (38.2, 38.8, 33.4, 39.0 and 29.9 g·L^–1) acetoin in five repeated cycles under the same conditions. This work therefore provided an efficient H. bluephagenesis whole-cell catalysis with a broad development prospect in biosynthesis of acetoin.     

Implications from γ-globulin adsorption onto cation exchangers fabricated by sequential alginate grafting and sulfonation

Our previous work proved that high adsorption capacity and uptake rate of lysozyme were achieved on alginate(Alg)-grafted re sin with an ionic capacity(IC) of 240 mmol·L~(-1)(Alg-FF-240).Moreover,the salt-tolerant feature of Alg-FF-230 was improved by using sequential alginate grafting and sulfonation strategy.Inspired by the enhanced adsorption performance of lysozyme,we have herein proposed to investigate the static and dynamic adsorption behaviors of γ-globulin on a series of Alg-grafted resins with different grafting densities and sulfonation degrees.The adsorption ca pacity of γ-globulin decreased with increa sing alginate-grafting density(IC) from 160 to 230 mmol·L~(-1) at 0 mmol·L~(-1) NaCl because of the steric hindrance caused by the alginate-grafting layer.Effects of ionic strength(IS) indicated that the adsorption capacities of the resins with the IC value of 230-370 mmol·L~(-1) were much higher than CM Sepharose FF at 50-100 mmol·L~(-1) NaCl,and the uptake rate of Alg-FF-230 was about twice as much as that of CM Sepharose FF.This work demonstrated the important effects of alginate-grafting layer and IS in γ-globulin adsorption behavior,which would be helpful in the design of Alggrafted resins and the selection of proper IS condition for protein purification.