马铃薯淀粉同步糖化发酵制备L-乳酸条件的统计学优化

以马铃薯淀粉为原料,采用同步糖化发酵方法制备乳酸。通过Plackett-Burman实验设计对影响乳酸产量的7个因子进行筛选,结果表明淀粉质量浓度、糖化酶用量和发酵温度3个因素对乳酸产量影响显著。利用最陡爬坡试验逼近最大响应区,采用中心复合实验设计及响应面分析法进行回归分析,建立影响乳酸产量的二次模型。模型求解得出最优淀粉质量浓度为271.89g/L,糖化酶用量为265.09U/g,发酵温度为39.05℃,最大理论乳酸产量为196.99g/L。3批验证实验结果平均值与预测值接近,表明该模型与实际情况拟合良好,实际最大乳酸产量为193.6g/L,较优化前提高了13.9%,L-乳酸的平均纯度达到95.2%。     

光合菌生物制氢技术

简要分析了光合细菌产氢的主要影响因素,介绍了国内外光合细菌生物制氢技术的研究和应用现状,并对光合制氢技术的发展趋势和应用前景进行了评述。     

Continuous fermentative hydrogen production from a wheat starch co-product by mixed microflora

For the transition to the hydrogen economy, hydrogen must be produced sustainably, e.g., by the fermentation of agricultural material. Continuous fermentative production of hydrogen from an insoluble substrate in nonsterile conditions is yet to be reported. In this study hydrogen production using mixed microflora from heat-treated digested sewage sludge in nonsterile conditions from a particulate co-product of the wheat flour industry (7.5 g L(-1) total hexose) at 18- and 12-hour hydraulic retention times, pH 4.5 and 5.2, 30 degrees C and 35 degrees C was examined. In continuous operation, hydrogen yields of approximately 1.3 moles hydrogen/mole hexose consumed were obtained, but decreased if acetate or propionate levels rose, indicating metabolism shifted towards hydrogen consumption by homoacetogenesis or propionate producers. These shifts occurred both at pH 4.5 and 5.2. Sparging the reactor with nitrogen to reduce hydrogen in the off-gas from 50% to 7% gave stable operation with a hydrogen yield of 1.9 moles hydrogen /mole hexose consumed over an 18-day period.     

固体发酵法固定工业废水处理优势菌的研究

为实现优势菌种的工程应用,由焦化废水处理站污泥筛选优势好氧菌A和兼性厌氧菌F,使用谷壳g、豆皮d为载体,采用固体发酵法进行固定化产品制备。初步研究表明固体发酵法固定菌种具有可行性,主要调控因素有温度、初始含水率、工业废水比例、pH、通风量和发酵时间。以pH8．0、温度好氧菌30℃（兼性厌氧菌35℃）、接种量1mL茴液儋田定化载体,初始含水量80％,固体发酵时间5d,低于60℃鼓风干燥,制备固定化产品。保存三个月,谷壳固定化产品好氧菌八的硝化能力和兼性厌氧菌R的反硝化能力不随保存时间的延长而改变;豆皮固定化产品氏、Fd降解能力随保存时问延长下降12％左右。以实验室岔／O工艺系统验证固定化产品处理焦化废水效果,缺氧池中无NO3-N积累,反硝化作用明显。氐和气对氰化物的去除率均达99％以上;对苯酚的去除效果气（98．84％）优于氏（79．6％）;但对NH4＋-N的去除率氏（75．46％）优于Ag（62．55％）。     

联合生物制氢方法及发展趋势

为了在生物制氢过程中最大限度提高产氢量和产氢速率,增大底物的利用率以及更好地发挥菌种间的协同作用,联合生物制氢技术成为近年来人们关注的焦点。综述了目前国内外几种联合生物制氢方法的研究现状。并从产氢机理的角度对几种联合制氢技术进行了分析比较,重点强调光合发酵和暗发酵联合生物制氢技术具有广泛的发展前景,并指出其存在的问题和未来的发展趋势。     

d- and l-lactic acid production from fresh sweet potato through simultaneous saccharification and fermentation

The aim of this study was to develop a bioprocess for l- and d-lactic acid production from raw sweet potato through simultaneous saccharification and fermentation by Lactobacillus paracasei and Lactobacillus coryniformis, respectively. The effects of enzyme and nitrogen source concentrations as well as of the ratio of raw material to medium were investigated. At dried material concentrations of 136.36–219.51 g L⁻¹, yields of 90.13–91.17% (w/w) and productivities of 3.41–3.83 g L⁻¹ h⁻¹ were obtained with lactic acid concentrations as high as 198.32 g L⁻¹ for l-lactic acid production. In addition, d-lactic acid was produced with yields of 90.11–84.92% (w/w) and productivities of 2.55–3.11 g L⁻¹ h⁻¹ with a maximum concentration of 186.40 g L⁻¹ at the same concentrations of dried material. The simple and efficient process described in this study will benefit the tuber and root-based lactic acid industries without requiring alterations in plant equipment.     

Enumeration of bacteria forming acetate from H2 and CO2 in anaerobic habitats

A method has been worked out that allows the detection and isolation of bacteria fermenting molecular hydrogen and carbon dioxide to acetic acid.The ratio of methanogenic to acetogenic bacteria in sludge and lake sediment samples has been found to be approximately 100 to 1. Acetogenic bacteria could not be detected in rumen samples.     

Aerobic submerged fermentation by acetic acid bacteria for vinegar production: Process and biotechnological aspects

Strictly aerobic acetic acid bacteria (AAB) have a long history of use in fermentation processes, and the conversion of ethanol to acetic acid for the production of vinegar is the most well-known application.At the industrial scale, vinegar is mainly produced by submerged fermentation, which refers to an aerobic process in which the ethanol in beverages such as spirits, wine or cider is oxidized to acetic acid by AAB. Submerged fermentation requires robust AAB strains that are able to oxidize ethanol under selective conditions to produce high-titer acetic acid. Currently submerged fermentation is conducted by unselected AAB cultures, which are derived from previous acetification stocks and maintained by repeated cultivation cycles.In this work, submerged fermentation for vinegar production is discussed with regard to advances in process optimization and parameters (oxygen availability, acetic acid content and temperature) that influence AAB activity. Furthermore, the potential impact arising from the use of selected AAB is described.Overcoming the acetification constraints is a main goal in order to facilitate innovation in submerged fermentation and to create new industry-challenging perspectives.     

Optimization of glutamate concentration and pH for H production from volatile fatty acids by Rhodopseudomonas capsulata

AIMS: This study attempted to employ response surface methodology (RSM) to evaluate the effects of glutamate concentration and pH on H(2) production from volatile fatty acids by Rhodopseudomonas capsulata. METHODS AND RESULTS: A mixture of acetate, propionate and butyrate was used as a carbon source for the H(2) production by R. capsulata. The H(2) yield and H(2) production rate were strongly affected by the glutamate concentration, pH and their interaction. The predicted maximum H(2) yield of 0.534 was obtained when glutamate concentration and pH were 6.56 mmol l(-1) and 7.29 respectively. On the contrary, the maximum H(2) production rate of 18.72 ml l(-1) h(-1) was achieved at a glutamate concentration of 7.01 mmol l(-1) and pH 7.31. CONCLUSIONS: Taking H(2) yield and H(2) production rate together into account, a glutamate concentration of 6.56-7.01 mmol l(-1) and pH of 7.29-7.31 should be selected for H(2) production from a mixture of acetate, propionate and butyrate by R. capsulata. SIGNIFICANCE AND IMPACT OF THE STUDY: The RSM was a useful tool for maximizing H(2) production by photosynthetic bacteria (PSB).     

Optimization of enzymatic hydrolysis and fermentation conditions for improved bioethanol production from potato peel residues

The aim of this work was the optimization of the enzyme hydrolysis of potato peel residues (PPR) for bioethanol production. The process included a pretreatment step followed by an enzyme hydrolysis using crude enzyme system composed of cellulase, amylase and hemicellulase, produced by a mixed culture of Aspergillus niger and Trichoderma reesei. Hydrothermal, alkali and acid pretreatments were considered with regards to the enhancement of enzyme hydrolysis of potato peel residues. The obtained results showed that hydrothermal pretreatment lead to a higher enzyme hydrolysis yield compared to both acid and alkali pretreatments. Enzyme hydrolysis was also optimized for parameters such as temperature, pH, substrate loading and surfactant loading using a response surface methodology. Under optimized conditions, 77 g L^?1 of reducing sugars were obtained. Yeast fermentation of the released reducing sugars led to an ethanol titer of 30 g L^?1 after supplementation of the culture medium with ammonium sulfate. Moreover, a comparative study between acid and enzyme hydrolysis of potato peel residues was investigated. Results showed that enzyme hydrolysis offers higher yield of bioethanol production than acid hydrolysis. These results highlight the potential of second generation bioethanol production from potato peel residues treated with onsite produced hydrolytic enzymes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:397–406, 2017     

泡菜中优良乳酸菌的分离、鉴定及发酵特性

从几种泡菜中分离出86株菌,对其在适温和低温下产酸速率及硝酸盐降解能力进行测定,筛选出5株产酸速率快、硝酸盐降解能力强的菌株。经形态学鉴定及生理生化反应试验,初步鉴定为：植物乳杆菌2株,短乳杆菌1株,戊糖乳杆菌1株,肠膜明串珠菌葡聚糖亚种1株,并对5株菌的发酵性能进行了测定。     

Application of simultaneous saccharification and fermentation (SSF) from viscosity reducing of raw sweet potato for bioethanol production at laboratory, pilot and industrial scales

The aim of this work was to research a bioprocess for bioethanol production from raw sweet potato by Saccharomyces cerevisiae at laboratory, pilot and industrial scales. The fermentation mode, inoculum size and pressure from different gases were determined in laboratory. The maximum ethanol concentration, average ethanol productivity rate and yield of ethanol after fermentation in laboratory scale (128.51 g/L, 4.76 g/L/h and 91.4%) were satisfactory with small decrease at pilot scale (109.06 g/L, 4.89 g/L/h and 91.24%) and industrial scale (97.94 g/L, 4.19 g/L/h and 91.27%). When scaled up, the viscosity caused resistance to fermentation parameters, 1.56 AUG/g (sweet potato mash) of xylanase decreased the viscosity from approximately 30000 to 500 cp. Overall, sweet potato is a attractive feedstock for be bioethanol production from both the economic standpoints and environmentally friendly.     

Interactions between Clostridium beijerinckii and Geobacter metallireducens in co‐culture fermentation with anthrahydroquinone‐2, 6‐disulfonate (AH2QDS) for enhanced biohydrogen production from xylose

To enhance biohydrogen production, Clostridium beijerinckii was co‐cultured with Geobacter metallireducens in the presence of the reduced extracellular electron shuttle anthrahydroquinone‐2, 6‐disulfonate (AH₂QDS). In the co‐culture system, increases of up to 52.3% for maximum cumulative hydrogen production, 38.4% for specific hydrogen production rate, 15.4% for substrate utilization rate, 39.0% for substrate utilization extent, and 34.8% for hydrogen molar yield in co‐culture fermentation were observed compared to a pure culture of C. beijerinckii without AH₂QDS. G. metallireducens grew in the co‐culture system, resulting in a decrease in acetate concentration under co‐culture conditions and a presumed regeneration of AH₂QDS from AQDS. These co‐culture results demonstrate metabolic crosstalk between the fermentative bacterium C. beijerinckii and the respiratory bacterium G. metallireducens and suggest a strategy for industrial biohydrogen production. Biotechnol. Bioeng. 2013; 110: 164–172. © 2012 Wiley Periodicals, Inc.     

Effect of initial bacteria concentration on hydrogen gas production from cheese whey powder solution by thermophilic dark fermentation

Dark fermentative hydrogen gas production from cheese whey powder solution was realized at 55°C. Experiments were performed at different initial biomass concentrations varying between 0.48 and 2.86 g L^?1 with a constant initial substrate concentration of 26 ± 2 g total sugar (TS) per liter. The highest cumulative hydrogen evolution (633 mL, 30°C), hydrogen yield (1.56 mol H₂ mol^?1 glucose), and H₂ formation rate (3.45 mL h^?1) were obtained with 1.92 g L^?1 biomass concentration. The specific H₂ production rate decreased with increasing biomasss concentration from the highest value (47.7 mL g^?1 h^?1) at 0.48 g L^?1 biomass concentration. Total volatile fatty acid concentration varied beetween 10 and 14 g L^?1 with the highest level of 14.2 g L^?1 at biomass concentration of 0.48 g L^?1 and initial TS content of 28.4 g L^?1. The experimental data were correlated with the Gompertz equation and the constants were determined. The most suitable initial biomass to substrate ratio yielding the highest H₂ yield and formation rate was 0.082 g biomass per gram of TS. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 28: 931–936, 2012     

不同生境微生物转化H2/CO2产乙酸及其在合成气发酵中应用

【目的】合成气发酵对大力开发可再生资源和促进国家可持续发展具有重要意义,研究旨在探究不同生境微生物转化H2/CO2产乙酸及其合成气发酵的潜力。【方法】采集剩余污泥、牛粪、产甲烷污泥和河道底物样品在中温(37 °C)条件下生物转化H2/CO2气体,将来源于牛粪样品的H2/CO2转化富集物用于合成气发酵,通过454高通量技术和定量PCR技术分析复杂微生物群落的组成,GC气相色谱法检测气体转化产生的挥发性脂肪酸(VFAs)浓度。【结果】牛粪和剩余污泥微生物利用H2/CO2气体生成乙酸、乙醇和丁酸等,最高乙酸浓度分别为63 mmol/L和40 mmol/L,明显高于河道底物和产甲烷污泥样品的最高乙酸浓度3 mmol/L和16 mmol/L。牛粪和剩余污泥微生物中含有种类多样化的同型产乙酸菌,剩余污泥中同型产乙酸菌主要为Clostridium spp.、Sporomusa malonica和Acetoanaerobium noterae,牛粪中则为Clostridium spp.、Treponema azotonutricium和Oxobacter pfennigii。【结论】同型产乙酸菌的丰富度和数量两个因素都对复杂微生物群落转化H2/CO2产乙酸效率至关重要;转化H2/CO2得到的富集物可用于合成气发酵产乙酸和乙醇,这为基于混合培养技术的合成气发酵提供了依据。     

The relationship between instability of H2 production and compositions of bacterial communities within a dark fermentation fluidized-bed bioreactor

Microbial community composition dynamics was studied during H(2) fermentation from glucose in a fluidized-bed bioreactor (FBR) aiming at obtaining insight into the H(2) fermentation microbiology and factors resulting in the instability of biofilm processes. FBR H(2) production performance was characterised by an instable pattern of prompt onset of H(2) production followed by rapid decrease. Gradual enrichment of organisms increased the diversity of FBR attached and suspended-growth phase bacterial communities during the operation. FBR bacteria included potential H(2) producers, H(2) consumers and neither H(2) producers nor consumers, and those distantly related to any known organisms. The prompt onset of H(2) production was due to rapid growth of Clostridium butyricum (99-100%) affiliated strains after starting continuous feed. The proportion trend of C. butyricum in FBR attached and suspended-growth phase communities coincided with H(2) and butyrate production. High glucose loading rate favoured the H(2) production by Escherichia coli (100%) affiliated strain. Decrease in H(2) production, associated with a shift from acetate-butyrate to acetate-propionate production, was due to changes in FBR attached and suspended-growth phase bacterial community compositions. During the shift, organisms, including potential propionate producers, were enriched in the communities while the proportion trend of C. butyricum decreased. We suggest that the instability of H(2) fermentation in biofilm reactors is due to enrichment and efficient adhesion of H(2) consumers on the carrier and, therefore, biofilm reactors may not favour mesophilic H(2) fermentation.     

H2 and acetate transfers during xylan fermentation between a butyrate-producing xylanolytic species and hydrogenotrophic microorganisms from the human gut

The aim of this work was to investigate in vitro interrelationships during xylan fermentation between an H2 and butyrate-producing xylanolytic species recently isolated in our laboratory from human faeces and identified as Roseburia intestinalis and the H2-utilizing acetogen Ruminococcus hydrogenotrophicus or the methanogen Methanobrevibacter smithii. H2 transfer between M. smithii or Ru. hydrogenotrophicus and the xylanolytic species was evidenced, confirming the great potential of these H2-consuming microorganisms to reutilize fermentative H2 during fibre fermentation in the gut. In addition, acetate transfer was demonstrated between the xylanolytic Roseburia sp. and the acetogenic species, both metabolites transfers leading to butyric fermentation of oat xylan without production of H2.     

水质净化乳酸菌的分离鉴定及发酵参数优化

【目的】从水产养殖环境和养殖生物体中选育具有水质净化功能的乳酸菌,以期为水产养殖提供专用高效的菌种资源。【方法】在低温和常温条件下从皮皮虾、南美白对虾肠道及养殖池底质活性污泥中分离具有水质净化功能的乳酸菌,对筛选的优良菌株采用形态、生理生化实验及16S r RNA序列分析进行鉴定,并对菌株的发酵参数进行了研究。【结果】低温和常温条件下从3种介质中共分离到乳酸菌136株,经水质净化能力筛选,发现常温分离的r13对模拟水体中亚硝态氮去除效果较强,72 h能将11.5 mg/L的亚硝态氮彻底去除,且对13.0 mg/L氨氮的去除率达到29.1%。经形态特征、生理生化特性及16S r RNA基因序列分析,鉴定菌株r13为植物乳杆菌Lactobacillus plantarum。发酵参数研究结果表明,该菌最适培养基组成为:酵母膏6.0 g/L,葡萄糖20.0 g/L,乙酸钠4.0 g/L,柠檬酸氢二铵2.0 g/L,K_2HPO_4 2.0 g/L,番茄汁50 m L/L;培养条件为:初始p H 6.0,接种量5%,装液量45/50,培养温度为34°C;在上述优化培养条件下发酵72 h,菌液的生物量达28.4 g/L湿重,有效活菌浓度达4.4×10~9 CFU/m L。     

Simultaneous removal of chemical oxygen demand (COD), phosphate, nitrate and H2S in the synthetic sewage wastewater using porous ceramic immobilized photosynthetic bacteria

Simultaneous aerobic treatment of COD, phosphate, nitrate and H₂S in a synthetic sewage wastewater was carried out using porous ceramic immobilized photosynthetic bacteria, Rhodobacter sphaeroidesS, Rb. sphaeroidesNR-3 and Rhodopseudomonas palustris. In the batch treatment, effective simultaneous removal of COD (89%), phosphate (77%), nitrate (99%) and H₂S (99.8%) was observed after 48 h. In semi-continuous treatments with dilution rates of 0.17 to 0.75 day^–1under aerobic conditions, simultaneous removal of these four components was also observed after about one month.     

Extractive fermentation for enhanced gellan-hydrolysing enzyme production by Bacillus thuringiensis H14

A new extractive fermentation process using PEG and potassium phosphate aqueous two-phase system (ATPS) was developed for enhanced production of gellan-hydrolysing enzyme by Bacillus thuringiensis H14. Five different Bacillus sp. were tested for their ability to synthesize gellan-hydrolysing enzyme. Bacillus thuringiensis H14 was found to be the best organism for gellan-hydrolysing enzyme production. The enzyme showed maximum activity at pH 7.5 and 40 °C. The partition studies of gellan-hydrolysing enzyme in the system using PEG X (X = 9000, 6000, 4000) and potassium phosphate–water and PEG–sodium citrate–water system indicated at PEG (4000)– potassium phosphate–water is the best system for partitioning of gellan-hydrolysing enzyme into the PEG phase (K = 4.99). Gellan-hydrolysing enzyme production by Bacillus thuringiensis H14 was studied in ATPSs composed of PEG X (X = 9000, 6000, 4000) and potassium phosphate. The top phase is continuous and rich in PEG while the bottom phase is dispersed and is rich in phosphate, microbial cells being mainly retained in the bottom phase. The gellan-hydrolysing enzyme produced during fermentation partitioned into the upper PEG phase and total gellan-hydrolysing enzyme produced was 2.12, 2.29 and 2.40 times higher than that of homogeneous fermentation when the fermentations were carried out using PEG 9000–potassium phosphate–water, PEG 6000–potassium phosphate–water, PEG 4000–potassium phosphate–water systems respectively.