豆粕水解液为氮源细菌厌氧流加发酵生产L-乳酸

采用细菌厌氧发酵法生产L-乳酸,由实验确定了最佳接种量、发酵温度和pH调节剂,考察了初始葡萄糖浓度对L-乳酸生产的影响,确定初始糖浓度为70~90 g/L时得率、产率、最终生物量分别达到92.68 g/g, 3.17 g/(L×h)和8.5′107 mL-1. 为进一步降低L-乳酸生产成本,以豆粕水解液为氮源代替酵母粉,同时应用流加发酵技术,L-乳酸产量、得率、产率及转化率分别达到155 g/L, 95.5 g/g, 1.64 g/(L×h)和96.9%. 在保证L-乳酸最终浓度的同时可降低生产成本,为进一步工业化奠定了基础.     

聚乙烯醇丝固定化米根霉发酵产L（+）-乳酸

以聚乙烯醇(PVA)丝为吸附材料的立体载体,固定化米根霉菌种As3.3462发酵生产乳酸。与游离发酵相比,摇瓶单批固定化发酵乳酸的最终质量浓度提高了25%,发酵时间缩短了33%。分别在摇瓶和鼓泡式反应器中进行了载体的稳定性研究,摇瓶中可稳定发酵10批次,平均转化率为65%,平均产率为1.1g/(L.h);1L鼓泡式生物反应器的放大实验中进行了9批半连续发酵实验,乳酸质量浓度最高达到55.68g/L,转化率69.60%。     

利用纤维原料在串联式生物反应器中协同酶解发酵乳酸

采用廉价的纤维素原料代替粮食发酵生产乳酸,具有重要的社会意义和经济价值。在串联式生物反应器中,将纤维原料酶水解与固定化乳酸杆菌发酵相耦联,纤维素的酶解产物葡萄糖被有效转化成乳酸,纤维素对乳酸的转化率和乳酸产率分别为70.3%和0.713g?(L?h)?1。在酶解体系中添加纤维二糖酶可以提高酶解得率。将酶解罐、固定化纤二糖酶柱和固定化细胞柱相串联,可有效消除纤维二糖积累所造成的反馈抑制作用,纤维素对乳酸的转化率和乳酸产率分别提高到90.6%和0.986g?(L?h)?1。串联式生物反应器性能稳定,在重复分批发酵工艺中,连续10批纤维素对乳酸的转化率平均为89.6%。采用分批添料工艺,纤维底物的终浓度可增加到200g?L?1,发酵终点的乳酸浓度达105.2g?L?1,乳酸产率为1.315g?(L?h)?1。对等量底物而言,反应时间明显缩短,同时纤维素酶的利用率也得到了有效提高。     

Effects of three main sugars in cane molasses on the production of butyric acid with <Emphasis Type="Italic">Clostridium tyrobutyricum</Emphasis>

The effects of three main sugars in cane molasses were investigated systematically to prepare a cost-effective medium for butyric acid bioproduction. Additionally, 30 g/L corn steep liquor was screened out as the suitable nitrogen source. In the batch fermentation of free cells, when 60 g/L glucose was the only carbon source, 21.28 g/L butyric acid was achieved after 30 h cultivation. Similar product concentration, productivity and yield were obtained when 60 g/L fructose was applied. The utilization of sucrose would bring about lower productivity (0.29 g/L·h) and product concentration (18.15 g/L), but the yield of butyric acid/sucrose (0.34 g/g) is almost the same as that from glucose or fructose (0.35 g/g). Finally, the sugar mixture (15 g/L glucose, 20 g/L fructose and 35 g/L sucrose) was employed to produce butyric acid in a fibrous-bed bioreactor (FBB), and 40.11 g/L butyric acid was produced with one simple fed-batch strategy.     

Fermentative production of L(+)‐lactic acid from starch hydrolyzate and corn steep liquor as inexpensive nutrients by batch culture of Enterococcus faecalis RKY1

BACKGROUND: Attempts were made to determine the lactic acid production efficiency of novel isolate, Enterococcus faecalis RKY1 using four different starches (corn, tapioca, potato, and wheat starch) with different concentrations (50, 75, 100, and 125 g L^?1) and corn steep liquor as an inexpensive nitrogen source. RESULTS: The yield of lactic acid from each starch was higher than 95% based on initial starch concentrations. High lactic acid concentration (129.9 g L^?1) and yield (1.04 g‐lactic acid g^?1‐starch) were achieved faster (84 h) from 125 g L^?1 of corn starch. Among the starches used, tapioca starch fermentation usually completed in a shorter incubation period. The final dry cell weight was highest (7.0 g L^?1) for the medium containing 75 g L^?1 of corn starch, which resulted in maximum volumetric productivity of lactic acid (3.6 g L^?1 h^?1). The addition of 30 g L^?1 corn steep liquor supplemented with a minimal amount of yeast extract supported both cell growth and lactic acid fermentation. CONCLUSION: Enterococcus faecalis RKY1 was found to be capable of growing well on inexpensive nutrients and producing maximum lactic acid from starches and corn steep liquor as lower‐cost raw materials than conventionally‐used refined sugars such as glucose, and yeast extract as an organic nitrogen source in laboratory‐scale studies. These fermentation characteristics are prerequisites for the industrial scale production of lactic acid. Copyright © 2008 Society of Chemical Industry     

大肠杆菌FMME-N-26生产琥珀酸的发酵条件优化和放大

琥珀酸(Succinic acid)被认为是白色生物技术生产的最具潜力的大宗化学品之一,在工业上具有广泛的应用。微生物发酵生产琥珀酸具有环境友好和可持续发展等优点,展现出良好的发展前景,但是存在得率低、副产物积累、生产强度低等问题。为了实现琥珀酸的高效生产,在3.6 L发酵罐中对E. coli FMME-N-26生产琥珀酸发酵条件和补料策略进行了优化,建立了好氧-厌氧两阶段发酵工艺,最终确定发酵策略为：有氧发酵8 h后转为厌氧发酵,MgCO3为pH中和剂,发酵72 h补加抗渗透压保护剂2 mmol/L甜菜碱,厌氧阶段控制葡萄糖浓度为1~5 g/L。优化后发酵72 h,琥珀酸的产量和厌氧阶段得率分别达到119.2 g/L和1.08 g/g葡萄糖(理论得率97%),分别比优化前提高了46.4%和4.8%,副产物乙酸和乳酸仅积累2.37和0.94 g/L,分别比优化前降低了37.1%和49.2%。在1000 L发酵罐中实现中试放大生产,E. coli FMME-N-26生产琥珀酸的产量、得率和生产强度在国内外属于领先水平,为琥珀酸工业化生产奠定了坚实的基础,同时也为其他高价值化学品的生产提供了借鉴。     

Detoxification and concentration of corn stover hydrolysate and its fermentation for ethanol production

Environmental and energy concerns have increased interest in renewable energy sources, particularly biofuels. Thus the fermentation of glucose from sulfuric acid-hydrolyzed corn stover for the production of bioethanol has been explored using a combined acid retardation and continuous-effect membrane distillation treatment process. This process resulted in the separation of the sugars and acids from the acid-catalyzed hydrolysate, the removal of most of the fermentation inhibitors from the hydrolysate and the concentration of the detoxified hydrolysate. The recovery rate of glucose from the sugar-acid mixture using acid retardation was greater than 99.12% and the sulfuric acid was completely recovered from the hydrolysate. When the treated com stover hydrolysate, containing 100 g/L glucose, was used as a carbon source, 43.06 g/L of ethanol was produced with a productivity of 1.79 g/(L· h) and a yield of 86.31 %, In the control experiment, where glucose was used as the carbon source these values were 1.97 g/(L·h) and 93.10% respectively. Thus the integration of acid retardation and a continuous-effect membrane distillation process are effective for the production of fuel ethanol from com stover.     

Use of sweet sorghum juice for lactic acid fermentation: preliminary steps in a process optimization

BACKGROUND: Lactic acid has many applications in the chemical industries and it can be produced economically by microorganisms using biomass raw materials of different origins. Sweet sorghum juice is a high sugar content raw material with potential for lactic acid production because after hydrolysis of its sucrose content the remaining glucose and fructose can supply the carbon demand of most lactic acid bacteria. However, satisfying the nitrogen and B‐vitamin needs of the bacteria by supplementation with yeast extract and/or other alternative nitrogen‐containing supplements can make the process too expensive. RESULTS: Using a statistical optimization process much of the yeast extract can be replaced by a cheaper alternative nitrogen source, namely wheat gluten. This resulted in a fermentation with 99% lactic acid yield and 3.04 g L^?1 h^?1 volumetric productivity. CONCLUSION: Using response surface methodology (RSM) media optimization was performed for lactic acid fermentation with an industrially acceptable result, reducing the costs of raw materials by half, replacing yeast extract by an alternative nitrogen source and applying yeast extract only as a source of micro‐elements (vitamins, salts, etc.) Copyright © 2010 Society of Chemical Industry     

铜离子与葡萄糖协同补料对球头三型孢菌产赤藓糖醇的影响

为考察葡萄糖和铜离子盐协同补料发酵对球头三型孢菌产赤藓糖醇的影响,在5L发酵罐中先采用不同浓度的葡萄糖进行分批发酵,然后采用优化的葡萄糖浓度并添加CuSO₄·5H₂O进行发酵研究。结果表明,初始葡萄糖浓度为300g/L的赤藓糖醇产量最大为44.52g/L,其体积生产速率为0.371g/（L·h）、转化率为0.167g/g。在此浓度葡萄糖的基础上添加30mg/L的CuSO₄·5H₂O后,赤藓糖醇产量达到49.62g/L,提高了11.5%。进一步控制总糖浓度为300g/L,且初始浓度为200g/L,分别进行单独补糖和协同补糖与铜离子的补料发酵,结果赤藓糖醇产量分别为47.25g/L和55.31g/L,比初始300g/L的葡萄糖分批发酵分别提高了6.1%和24.2%。特别地,协同补糖与CuSO₄·5H₂O后,赤藓糖还原酶（erythrose reductase,ER）的活性在84h达到最大,为0.152U/mg,比单独补糖时提高了18.8%;通过铜离子盐和葡萄糖的协同补料发酵可显著提高赤藓糖醇的产量,最终使赤藓糖醇产率达到0.461g/（L·h）。     

Production of bioethanol under high pressure of CO2: The effect of process conditions

The aim of this work is to investigate the possibility of producing ethanol by glucose fermentation under high pressure of carbon dioxide up to 48 bar, in order to exploit both ethanol and denser CO₂ as a by-product of the process. The fermentation is carried out using Ethanol Red™-Lesaffre Saccharomyces cerevisiae yeast strain, which is commonly applied in industrial bioethanol production.The experiments were performed in six small reactors (2 mL of volume each) connected in parallel, to investigate the effect of the process variables at the same conditions of temperature and pressure, and in one pilot reactor (1 L of volume) to confirm the results obtained at the lower scale.The influence of operative variables, such as carbon dioxide pressure (0–48 bar), temperature (32 and 36 °C), glucose (150–250 g/L), inositol (0–400 mg/L) and biomass concentration (OD 2 and 3.5), was measured in terms of ethanol concentration (by gas chromatography) and ethanol productivity (expressed as grams of ethanol per CFU of yeast). Both of these parameters were found to be strongly dependent on glucose concentration and CO₂ pressure, which negatively affects the fermentation. Nevertheless, also at 50 bar it is possible to produce appreciable amounts of ethanol.     

Bioethanol production from corn meal by simultaneous enzymatic saccharification and fermentation with immobilized cells of Saccharomyces cerevisiae var. ellipsoideus

The simultaneous enzymatic saccharification and fermentation (SSF) of corn meal using immobilized cells of Saccharomycescerevisiae var. ellipsoideus yeast in a batch system was studied. The yeast cells were immobilized in Ca-alginate by electrostatic droplet generation method. The process kinetics was assessed and determined and the effect of addition of various yeast activators (mineral salts: ZnSO₄ · 7H₂O and MgSO₄ · 7H₂O, and vitamins: Ca-pantothenate, biotin and myo-inositol) separately or mixed, was investigated. Taking into account high values of process parameters (such as ethanol concentration, ethanol yield, percentage of the theoretical ethanol yield, volumetric productivity and utilized glucose) and significant energy savings the SSF process was found to be superior compared to the SHF process. Further improvement in ethanol production was accomplished with the addition of mineral salts as yeast activators which contributed to the highest increase in ethanol production. In this case, the ethanol concentration of 10.23% (w/w), percentage of the theoretical ethanol yield of 98.08%, the ethanol yield of 0.55 g/g and the volumetric productivity of 2.13 g/l·h were obtained.     

Experimental evaluation of alternative fermentation media for L‐lactic acid production from apple pomace

BACKGROUND: A variety of nitrogen sources were tested for lactic acid production. Corn steep liquor is a low‐cost by‐product that could replace some of the expensive nutrients of the general lactobacilli media. This work deals with the optimisation of the composition of a low‐cost medium for lactic acid production from apple pomace by Lactobacillus rhamnosus CECT‐288. RESULTS: Corn steep liquor (CSL) and yeast extract (YE) were evaluated as nutrient sources for lactic acid production from apple pomace. In comparison with media containing CSL, experiments with YE led to higher volumetric productivities but lower lactic acid concentrations and product yields. The presence of YE increased the production of acetic acid. In media containing 0.2 g CSL g^?1 potential sugars, 29.5 g lactic acid L^?1 was obtained after 24 h, at good yield (35.5 g per 100 g dry apple pomace) and productivity (1.23 g L^?1 h^?1), with a lactic acid/acetic acid mass ratio of 98 g g^?1. CONCLUSION: The experimental results proved that CSL is a suitable nutrient source for lactic acid production from apple pomace, even though the volumetric productivity was lower than in experiments employing YE. Considered as a nutrient supplement, CSL presents advantages over YE not only in terms of cost but also in terms of product yield, final lactic acid concentration and lactic acid/acetic acid mass ratio. The utilisation of apple pomace as the raw material and CSL as the sole nutrient source (both cheap by‐products) enables the production of lactic acid by an economical, environmentally friendly process. Copyright © 2008 Society of Chemical Industry     

利用改性载体固定化大肠杆菌产琥珀酸

采用聚乙烯亚胺（PEI）和戊二醛（GA）对棉纤维进行化学修饰,考察了载体改性后的性能和对固定化大肠杆菌产丁二酸的影响。改性后的载体菌体负载量提高了63.3%。培养基中葡萄糖浓度为43 g/L,添加改性棉纤维120g/L,以MgCO3为缓冲盐,进行批式发酵,丁二酸浓度达到29.6 g/L,比未改性棉纤维提高了11.3%;丁二酸收率达到70.5%,比改性前提高了7.5%;丁二酸生产速率达到0.66 g/(L?h), 比改性前提高了37.5% 。对该材料固载的细胞进行7次重复批式发酵,丁二酸产量、转化率和产率没有下降趋势,具有一定的重复稳定性。     

代谢工程改造大肠杆菌生产琥珀酸

琥珀酸（succinic acid）是一种四碳二羧酸,在食品、医药、塑料和化工行业具有广泛的应用。目前,微生物法生产琥珀酸存在得率低、生产强度低、副产物积累等问题。为此,本研究通过复合诱变（ARTP和⁶⁰Co-γ射线）筛选到一株耐高渗突变株FMME-N-2,其琥珀酸得率为0.70g/g葡萄糖,同时积累18.8g/L乳酸、7.6g/L甲酸和17.3g/L乙酸。为了提高琥珀酸得率,通过敲除乳酸脱氢酶基因（ldhA）、丙酮酸-甲酸裂解酶-甲酸转运蛋白基因（pflB-focA）、磷酸转乙酰基基因（pta）、丙酸激酶基因（tdcD）和a-酮丁酸甲酸酯裂解酶基因（tdcE）,阻断冗余代谢支路减少副产物积累,获得工程菌株FMME-N-13,琥珀酸得率增加到0.92g/g葡萄糖,同时副产物大大降低,积累0.6g/L乳酸、3.6g/L甲酸和12.3g/L乙酸。同时,通过调控RBS强度组合优化来自产琥珀酸放线杆菌的磷酸烯醇式丙酮酸羧激酶基因（AsPCK）和来自博伊丁假丝酵母的甲酸脱氢酶基因（CbFDH）的表达水平,调控胞内ATP和NADH的浓度,最优工程菌FMME-N-26（FMME-N-13-L-AsPCK-L-CbFDH）的琥珀酸得率增加至1.04g/g葡萄糖,仅积累5.5g/L乙酸;最终,对厌氧阶段葡萄糖浓度进行优化,当葡萄糖浓度控制在0~5g/L时,菌株FMME-N-26的琥珀酸浓度增加到111.9g/L,得率为1.11g/g葡萄糖（理论产率的99%）,生产强度为1.76g/L/h,为琥珀酸的工业化生产奠定了良好的基础。     

Bioconversion of corn stover hydrolysate to ethanol by a recombinant yeast strain

Three corn stover hydrolysates, enzymatic hydrolysates prepared from acid and alkaline pretreatments separately and hemicellulosic hydrolysate prepared from acid pretreatment, were evaluated in composition and fermentability. For enzymatic hydrolysate from alkaline pretreatment, ethanol yield on fermentable sugars and fermentation efficiency reached highest among the three hydrolysates; meanwhile, ethanol yield on dry corn stover reached 0.175 g/g, higher than the sum of those of two hydrolysates from acid pretreatment. Fermentation process of the enzymatic hydrolysate from alkaline pretreatment was further investigated using free and immobilized cells of recombinant Saccharomyces cerevisiae ZU-10. Concentrated hydrolysate containing 66.9 g/L glucose and 32.1 g/L xylose was utilized. In the fermentation with free cells, 41.2 g/L ethanol was obtained within 72 h with an ethanol yield on fermentable sugars of 0.416 g/g. Immobilized cells greatly enhanced the ethanol productivity, while the ethanol yield on fermentable sugars of 0.411 g/g could still be reached. Repeated batch fermentation with immobilized cells was further attempted up to six batches. The ethanol yield on fermentable sugars maintained above 0.403 g/g with all glucose and more than 92.83% xylose utilized in each batch. These results demonstrate the feasibility and efficiency of ethanol production from corn stover hydrolysates.     

利用甘蔗糖蜜与乳清粉厌氧发酵制备丁二酸

考察了甘蔗糖蜜替代昂贵葡萄糖作为碳源、乳清粉替代大部分酵母粉作为氮源时,对Actinobacillus succinogenes NJ113发酵制备丁二酸的影响。血清瓶厌氧发酵结果证明：对照组（葡萄糖40 g/L）的丁二酸产量仅为26.04 g/L,而以糖蜜为碳源（以总还原糖计算为40 g/L）时,丁二酸产量达到28.27 g/L,比对照组提高了8.57%。在此基础上,以糖蜜为碳源、不同比例的乳清粉和酵母粉为混合氮源发酵制备丁二酸,确定了糖蜜、乳清粉和酵母粉混合使用的最佳浓度分别为40 g/L、8 g/L和2 g/L。此外,在3 L发酵罐体系中添加40 g/L糖蜜、8 g/L乳清粉、2 g/L酵母粉进行发酵试验,实验结果证明：丁二酸终产量达到32.54 g/L,收率达到81.13%。     

Strain isolation and optimization of process parameters for bioconversion of glycerol to lactic acid

BACKGROUND: The crude glycerol from biodiesel production represents an abundant and inexpensive source which can be used as raw material for lactic acid production. The first aim of this investigation was to select a strain suitable for producing lactic acid from glycerol with a high concentration and productivity. The second aim was to obtain the optimum fermentation conditions, as a basis for large‐scale lactate production in the future. RESULTS: Eight bacterial strains, which could aerobically convert glycerol to lactic acid, were screened from soil samples. One of the strains, AC‐521, which synthesized lactic acid with a higher concentration, was identified based on its 16S rDNA sequences and physiological characteristics. These results indicated that this strain was a member of Escherichia coli. The optimal fermentation conditions for Escherichia coli AC‐521 were 42 °C, pH 6.5, 0.85 min^?1 (K_La). CONCLUSION: Escherichia coli AC‐521 suitable for producing lactic acid from glycerol with high concentration and productivity was identified. After 88 h of fed‐batch fermentation, both the lactic acid concentration and glycerol consumption reached maximum, giving 85.8 g L^?1 of lactic acid with a productivity of 0.97 g L^?1 h^?1 and a yield of 0.9 mol mol^?1 glycerol. Copyright © 2009 Society of Chemical Industry     

Optimisation of a continuous flash fermentation for butanol production using the response surface methodology

Factorial design and response surface techniques were used in combination with mathematical modelling and computational simulation to optimise an innovative industrial bioprocess, the production of biobutanol employing the flash fermentation technology. A parametric analysis performed by means of a full factorial design at two levels determined the influence of operating variables on butanol yield and productivity. A second set of simulations were carried out based on the central composite rotatable design. This procedure generated simplified statistical models that describe butanol yield and productivity as functions of the significant operating variables. From these models, response surfaces were obtained and used to optimise the process. For a range of substrate concentration from 130 to 180 g/l, the optimum operating ranges ensure butanol productivity between 7.0 and 8.0 g/l h, butanol yield between 19 and 22%, substrate conversion above 90% and final butanol concentration around 25 g/l.     

兽疫链球菌NW-162发酵生产透明质酸的研究

在摇瓶中进行了兽疫链球菌诱变株NW-162发酵生产透明质酸的工艺研究,通过正交实验优化了最佳培养基组成,并考察了发酵过程中发酵温度、培养时间、pH值、装料系数等条件对该菌株发酵生产透明质酸的影响。实验表明:碳源及氮源对发酵过程影响最显著,优化所得最佳培养基组成为:葡萄糖40 g/L,复合氮源20 g/L,MgSO42g/L,钠盐3 g/L;在36℃、pH值7.0、装料系数为0.4的工艺条件下,发酵至27—30 h时达到最优发酵效果,收率可达0.464 g/L,比优化前提高2倍。     

酿酒酵母工程菌发酵产青蒿酸的工艺优化

通过酿酒酵母工程菌(Saccharomyces cerevisiae 1211)发酵制备青蒿酸,并通过单因素实验和响应面优化,考察了发酵温度、pH、半乳糖质量浓度、发酵碳源及发酵氮源等对青蒿酸发酵产量的影响。结果表明:在发酵温度30℃,发酵培养基初始pH=5.5,发酵培养基中蔗糖质量浓度91.8 g/L,半乳糖质量浓度10.1 g/L,硫酸铵质量浓度10.3 g/L,磷酸二氢钾质量浓度8.7 g/L的条件下,青蒿酸发酵产量可达(1529.7±12.6)mg/L,与未优化时的发酵产量相比,提升了67.1%。