发酵粗甘油产乳酸的戊糖乳杆菌代谢进化

以生物柴油产业的副产物粗甘油为底物,可降低乳酸发酵的生产成本。但是,粗甘油发酵生产乳酸存在菌体生长缓慢、菌浓较低、产酸速率和终产物浓度偏低等问题。以实验室筛选的一株戊糖乳杆菌 (Lactobacillus pentosus R3-8)为出发菌株进行代谢进化。通过在培养基中添加高浓度的粗甘油和乳酸,分别进行菌株耐底物和产物抑制的代谢进化。用粗甘油驯化的第60代菌株,可耐受130 g·L^-1的粗甘油,与出发菌株相比,生长速率提高, 且生物量是原始菌株的1.23倍。用乳酸驯化的第50代菌株可耐受20 g·L^-1的乳酸,生物量比初始菌株提升了18%。驯化菌株的5 L发酵罐分批发酵结果显示,以粗甘油驯化至 60 代的菌株的批次发酵水平相对较好,乳酸产量、甘油转化率以及生产强度分别为 45.0 g·L^-1、0.989 g·g^-1和 0.47 g·L^-1·h^-1。以粗甘油驯化至 60 代的菌株进行补料分批发酵,乳酸终浓度为83.8 g·L^-1,比分批发酵提高了近1倍。     

FED-BATCH SIMULTANEOUS SACCHARIFICATION AND FERMENTATION OF MICROWAVE/ACID/ALKALI/H2O2 PRETREATED RICE STRAW FOR PRODUCTION OF ETHANOL

The batch simultaneous saccharification and fermentation (SSF) of microwave/acid/alkali/H₂O₂ pretreated rice straw to ethanol was optimized using cellulase from Trichoderma reesei and Saccharomyces cerevisiae YC-097 cells prior to the fed-batch SSF studies. The batch SSF optima were 10% w/v substrate, 40°C, 15 mg cellulase/g substrate, initial pH 5.3, and 72 hours. Under the optimum conditions the ethanol concentration and its yield were 29.1 g/L and 61.3% respectively. Based on the optimal batch SSF, the fed-batch SSF was investigated and its operation parameters were optimized. Under its optimal conditions the ethanol concentration reached 57.3 g/L, while its productivity and yield were only slightly less than those in the batch SSF. This suggests that fed-batch SSF is a potential operation mode for effective ethanol production from microwave/acid/alkali/H₂O₂ pretreated rice straw.     

Batch and Continuous Production of Lactic Acid from Beet Molasses by Lactobacillus delbrueckii IFO 3202

Process variables were optimized for the production of lactic acid from pretreated beet molasses by Lactobacillus delbrueckii IFO 3202 for batch and continuous fermentations. In the batch fermentation, maximum yields (95·4% conversion, 77·1% effective) and maximum lactic acid volumetric productivity (4·83 g dm⁻³ h⁻¹) was achieved at 45°C, pH 6·0, 78·2 g dm⁻³ sugar concentration with 10 g dm⁻³ yeast extract. Various cheaper nitrogen sources were replaced with yeast extract on equal nitrogen bases in batch fermentation. Of all the nitrogen sources tested, yeast extract yielded the highest and malt sprouts yielded the second highest level of lactic acid. In the continuous fermentation, maximum lactic acid (4·15%) was obtained at a dilution rate of 0·1 h⁻¹. Maximum volumetric lactic acid productivity (11·20 g dm⁻³ h⁻¹) occurred at D = 0·5 h⁻¹ dilution rate. © 1997 SCI.     

双极膜电渗析法分离发酵液中乳酸及离子迁移规律

考察了双极膜电渗析法分离酒糟发酵液中乳酸的影响因素,结果表明:当进料浓度13.5 g·L^-1,工作电压27 V,进料浓度比1.3,流量20 L·h^-1,电渗析10 h,产品罐中的乳酸浓度增加至20.95 g·L^-1,是电渗析前进料罐中的乳酸浓度的1.6倍,具有显著的浓缩效果.酸根离子迁移规律的研究表明,乳酸根离子的迁移速率明显高于其他酸根离子,且丙酸和乙酸的乳酸分离率相对较低.     

生物基1， 3-丙二醇连续发酵过程的稳态分析与反馈控制

生物发酵甘油生产1,3-丙二醇的自动化控制是其工业化应用亟待解决的关键问题。首先,采用数学函数连续性分析深入研究了克雷伯氏杆菌连续发酵甘油生产1,3-丙二醇过程的多稳态特性。在不同的初始甘油浓度或稀释速率下,系统均会出现多稳态现象,通过双因素分析确定了多稳态出现的临界区域,该区域内部的稳态是不稳定的。之后,基于反馈控制理论和多稳态分析结果,设计优化了受残余甘油和产物浓度影响的稀释速率控制策略。在连续发酵过程中,调整时间从81.27 h缩短到34.11 h,显著提高了发酵初期阶段甘油的利用率;同时甘油转化率由0.478 mmol·mmol^-1提高至0.563 mmol·mmol^-1,生产强度由85.70 mmol·L^-1·h^-1提高到101.10 mmol·L^-1·h^-1,显著提高了生产性能。     

REACTION KINETICS PARAMETERS DESCRIBING DISPLACEMENT BY ACIDS OF STRUCTURAL METALLIC IONS FROM CLAY MINERALS

Chemical kinetics parameters describing reactions of naturally-occurring clay minerals with acids were determined using batch reaction and analytical techniques. Clay minerals included sodium montmo-rillonite, kaolinite, and illite, reacting with hydrochloric acid, nitric acid, and sulfuric acid. Two acid concentrations were examined at 50 and 70^°C. Displacement by acids of aluminum and magnesium from clays was found to follow pseudo-first order kinetics. Data analysis verified the batch reaction technique for determining kinetics parameters of acid-clay reactions and demonstrated acid neutralization by naturally-occurring clays. For reactions of sodium montmorillonite with hydrochloric acid, nitric acid, and sulfuric acid, reaction rate coefficients for displacement of aluminum ranged from 1·73 × 10^-4 kg/mol-hr at 50^°C to 3·02 × 10^-3 kg/mol-hr at 70^°C. Displacement of magnesium from sodium montmorillonite by these acids occurred at rates ranging from 3·68 × 10^-4 kg/mol-hr at 50^°C to 1·12 × 10^-2 kg/mol-hr at 70^°C. Activation energies for displacement by acids of aluminum from sodium montmorillonite were calculated to range from 31,000J/mol to 112,000J/mol, while activalion energies for displacement of magnesium from sodium montmorillonite ranged from 36.000 J/mol to 103,000 J/mol. For reactions of illite with acids, reaction rate coefficients for displacement of aluminum were determined to be 1·45×10^-4Kg/mol-hr at 50^°C to 1·7×10^-3kg/mol-hr at 70^°C. Displacement of magneisum by acids from illite occurred at rates from 6·06 × 10^-4 kg/mol-hr at 50^°C to 5·97 × 10^-3 kg/mol-hr. Activation energies for reactions of acids with illite ranged from 33,000 J/mol to 62,000 J/mol for displacement of aluminum and from 9,400 J/mol to 29,000 J/mol for displacement of magnesium.     

Batch kinetics and modelling of propionic acid fermentation

Batch propionic acid fermentation kinetics was studied using five different initial concentrations of lactose (i.e., 37 g/L, 45g/L, 50g/L, 57 g/L and 73 g/L) at constant temperature (30°C) and pH (6.5) under anaerobic conditions using Propionibacterium acidipropionici (ATCC 4875). When the initial substrate concentration was 37 g/L, 45 g/L, 50 g/L, 57 g/L and 73 g/L, then, correspondingly, 16 g/L, 19 g/L, 22.25 g/L, 25.3 g/L and 26.3 g/L of propionic acid was accumulated in the fermentation broth. Increasing the supply of lactose in the fermentation medium led to the accumulation of by products succinate, acetate and pyruvate. Maximum propionate yield (0.44 g/g) and comparatively lesser impurities (byproducts) were achieved with 57 g/L initial lactose concentration. The batch growth kinetics was eventually used to develop and test a mathematical model for propionic acid fermentation by P. acidipropionici at pH 6.5 and S_o = 57 g/L. Y_X/S^max was found to be the most sensitive parameter of the model. The same model also successfully simulated the batch kinetics observed at S_o = 37 g/L. However the model failed to simulate the fermentation kinetics observed at S_o = 73 g/L. The developed model can be used for process optimization studies.     

Enhanced production of l(+)-lactic acid by floc-form culture of Rhizopus oryzae

A process to optimize l-lactic acid production from glucose by Rhizopus oryzae, based on sustaining floc morphology throughout the fermentation process, is herein performed. During the fermentation, supplementary ammonium sulfate was added intermittently to maintain the ammonia level of the culture medium always higher than 0.1 g/L. With replenish of nitrogen source, mycelia flocs did not aggregate, and the lactic acid production was optimized upon the fermentation being controlled at pH 4.3–4.5 by adding calcium carbonate slurry. In contrast, without supplementary addition of nitrogen source, mycelial clumps formed, resulting in a poor production of lactic acid. In the initial batch fermentation process, the final concentration of lactic acid produced was 109 g/L, with a yield (g lactic acid/g glucose consumed) of 0.87 and a productivity of 2.73 g/L h, using 125 g/L of glucose as substrate. For the first four cycles of repeated-batch fermentation, the average final concentration, the productivity and the yield of lactic acid were 113 g/L, 4.03 g/L h and 0.90, respectively.     

Further studies on redoxokinetic titrations

The previously described “redoxokinetic effect” is used to indicate the end-points of titrations of Fe²⁺ with Cr₂O₇^2-, sulphuric acid with sodium hydroxide, AsO₂- with I₂ and Ag⁺ with Cl⁻. With the first three systems an accuracy of 0·1 per cent is possible. The method is not suited to the fourth system.

Abstract

Titrations of sulphuric acid vs. sodium hydroxide at 0·1 N concentration and ferrous ammonium sulphate vs. potassium dichromate at 0·05 N concentration can be carried out with an accuracy of ±0·1 per cent using the redoxokinetic technique. A very sharp end-point was obtained in the case of iodine vs. arsenite titration at 0·1 N concentration. Silver nitrate vs. chloride titrations cannot be carried out by the redoxokinetic technique.

Addition of MnSO₄ to the extent of 50 g/l. of the solution enhances the precision considerably in the titration of dilute solutions of ferrous ammonium sulphate with dichromate.     

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 lactic acid from beet molasses by calcium alginate immobilized Lactobacillus delbrueckii IFO 3202

Lactic acid was produced from pretreated beet molasses by the homofermentative organism Lactobacillus delbrueckii subsp delbrueckii IFO 3202 entrapped in calcium alginate gel using batch, repeated batch and continuous fermentation systems. In batch fermentation studies successful results were obtained with 2.0–2.4 mm diameter beads prepared from 2% sodium alginate solution. The highest effective yield (82.0%) and conversion yield (90.0%) were obtained from substrate concentrations of 52.1 and 78.2 g dm⁻³ respectively. The gel beads produced lactic acid for 14 consecutive batch fermentations without marked activity loss and deformation. In the continuous fermentation, the highest lactic acid (4.22%) was obtained at a dilution rate of 0.1 h⁻¹ while the highest productivity (13.92 g dm⁻³ h⁻¹) was obtained at a dilution rate of 0.4 h⁻¹. © 1999 Society of Chemical Industry     

PRODUCT INHIBITION OF WHEY LACTOSE HYDROLYSIS

In the present study, glucose and galactose inhibition effects for β-galactosidase hydrolyzing lactose recovered from whey were investigated. The experiments were carried out in 250 mL of 25 mM phosphate buffer solution containing 1, 1.5, 2, 4, and 6% (w/v) lactose recovered from whey by using a commercial β-galactosidase produced from Kluyveromyces marxianus lactis at a constant temperature of 37°C, pH 6.5, and enzyme concentration of 1 mL/L, in a batch reactor system. The amounts of glucose and galactose added to the reaction solution were 6.25, 12.5, and 25 g/L. A second-order kinetic expression effectively simulated the data of residual lactose concentration with respect to processing time for each experimental condition examined. The Lineweaver-Burk plots showed that the inhibition effects of glucose and galactose were uncompetitive. The inhibition constants (K_i) obtained for glucose and galactose (10.32 g/L and 13.03 g/L, respectively) showed that the glucose was the most effective inhibitor for β-galactosidase.     

OPTIMIZATION OF PHOTOCATALYTIC PROCESS PARAMETERS FOR THE DEGRADATION OF 2,4,6-TRICHLOROPHENOL IN AQUEOUS SOLUTIONS

In the present study, the photocatalytic degradation of 2,4,6-trichlorophenol (2,4,6-TCP) was carried out in a batch reactor under UV light in aqueous solution for 5 h using titania P-25 (surface area 50 m²/g) as a photocatalyst and sodium hypochlorite as an oxidant. Variables studied include catalyst dose (0.25-1.25 g/L), pH (2-6), and concentration of the oxidant (3.06 × 10^-6M-15.3 × 10^-6M). The rate of degradation was studied in terms of changes in concentration of the pollutant and reduction in chemical oxygen demand. The optimal values of operational parameters leading to 2,4,6-TCP abatement were obtained by using response surface methodology. The percent degradation and COD reduction of 2,4,6-TCP was found to increase with increases in the catalyst dose up to the dose of 1.1 g/L, pH in the range of 4-4.5, and oxidant concentration of 9.95 × 10^-6M.     

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。     

CORRELATION OF GLUCOSE (DEXTROSE) DEGRADATION AT 90 TO 190°C IN 0.4 TO 20% ACID

Decomposition of glucose is described over a wide range of acid concentrations and temperatures by a correlation based on an Arrhenius type model combined with a modified Debye-Huckel equation. Degradation depends on temperature and acid concentration, represented by hydrogen ion activity. Kinetic data from our laboratory for 4 and 12 wt% glucose were combined with literature data to establish this model. Alternative non-linear mechanisms were compared by regression analysis of the published data as well as data developed in this study. Values of the degradation constant vary from 0.109 hr^-1 (at 20 wt% acid and 190°C) to 4.18 × 10^-7 (at 0.4 wt% acid and 100°C). This result will be useful to model glucose degradation in acid hydrolysis of celluloses, sterilization of fermentor feeds, and stripping of fermentation ethanol.     

MASS TRANSFER IN BANANA CHIPS DURING OSMOTIC DEHYDRATION

The effects of temperature ( 50, 60 and 70 °C ), sucrose concentration ( 50, 60 and 70° Brix ) and pH ( 6,7 and 8 ) on the mass transfer during osmotic dehydration of banana chips were studied. Fitting a diffusion model gave apparent diffusivity values varied from 2.77 to 2.66 × 10^-9 m² s^-1 and depending on temperature and sucrose concentration but not on pH changes. The effct of syrup/product volume ratio on the mass transfer kinetics was also investigated.     

Purification and concentration of gluconic acid from an integrated fermentation and membrane process using response surface optimized conditions

A response surface method was used to optimize the purification and concentration of gluconic acid from fermentation broth using an integrated membrane system. Gluconobacter oxydans was used for the bioconversion of the glucose in sugarcane juice to gluconic acid (concentration 45 g·L^-1) with a yield of 0.9 g·g^-1. The optimum operating conditions, such as trans-membrane pressure (TMP), pH, cross-flow rate (CFR) and initial gluconic acid concentration, were determined using response surface methodology. Five different types of polyamide nanofiltration membranes were screened and the best performing one was then used for downstream purification of gluconic acid in a flat sheet cross-flow membrane module. Under the optimum conditions (TMP=12 bar and CFR=400 L·h^-1), this membrane retained more than 85% of the unconverted glucose from the fermentation broth and had a gluconic acid permeation rate of 88% with a flux of 161 L·m^-2·h^-1. Using response surface methods to optimize this green nanofiltration process is an effective way of controlling the production of gluconic acid so that an efficient separation with high flux is obtained.     

Kinetics of Lactic Acid Fermentation by Lactobacillus delbrueckii Grown on Beet Molasses

The fermentation kinetics for the conversion of beet molasses, a valuable and economical fermentation substrate, to lactic acid by the homofermentative organism Lactobacillus delbrueckii C.E.C.T. 286 have been studied at controlled pH and temperature under anaerobic batch conditions. An inhibitory effect of lactic acid on fermentation of beet molasses has been found. The bacterium was able to produce lactic acid even after growth ceased. A kinetic model for the fermentation is proposed. From this model, the maximum allowable lactic acid concentration above which growth stops and the lactic acid level above which bacteria stop producing lactic acid were found to be 45 g dm⁻³ and 57 g dm⁻³, respectively. © 1997 SCI.     

PRODUCTION OF ACETONE AND BUTANOL BY CLOSTRIDIUM ACETOBUTYLICUM IN A PRODUCT LIMITED CHEMOSTAT

The solvents acetone and butanol are produced by Clostridium acetobutylicum during batch culture only after a considerable lag. The influence of limiting nutrients on the onset of solvent production has been investigated by studying the continuous fermentation under conditions of carbon, nitrogen and phosphate excess over a wide range of dilution rates. A maximum solvent concentration of 12 gl^-1 was obtained at the lowest dilution rate investigated, namely 0.05 hr^-1, and the average yield of solvents over the range of dilution rates approximated 0.3. These data confirm that a limitation of either of these nutrients is not essential for solvent production.     

A comparison of the oleaginous yeast,Candida curvata,grown on different carbon sources in continuous and batch culture

The oleaginous yeast,Candida curvata D, was grown in both batch and continuous culture on 5 different carbon sources to compare the efficiency of fat production from the various substrates. Maximum lipid accumulation occurred in batch culture with xylose as the carbon source on nitrogenlimited medium reaching a level of 49% (w/w) of the biomass, but this was reduced to 37% at the optimum dilution rate (D=0.05/hr) in a chemostat. Both the highest biomass and lipid yields were attained in continuous culture with lactose as the sole carbon source at a dilution rate of D=0.04/hr, giving an efficiency of substrate conversion of 60 g of biomass and 18.6 g lipid per 100 g lactose utilized. The relative proportions of the major fatty acids (16∶0, 18∶0, 18∶1, 18∶2) in the lipid were found to vary considerably in batch culture and in continuous culture under carbon-limited conditions. However, on nitrogen-limited media in the chemostat, the fatty acid composition remained relatively constant over the whole range of dilution rates employed. Lipid from xylose-grown cells contained the greatest percentage of stearic acid (18∶0) 15% and the lowest linoleic acid (18∶2) 4%, whereas lipid from ethanol-grown cells contained elevated levels of oleic acid (18∶1) 51% and decreased palmitic acid (16∶0) 25%.