l-Lactic acid production from liquefied cassava starch by thermotolerant Rhizopus microsporus: Characterization and optimization

The thermotolerant Rhizopus microsporus DMKU 33 capable of producing l-lactic acid from liquefied cassava starch was isolated and characterized for its phylogenetic relationship and growth temperature and pH ranges. The concentrations of (NH₄)₂SO_4, KH₂PO₄, MgSO₄ and ZnSO₄·7H₂O in the fermentation medium was optimized for lactic acid production from liquefied cassava starch by Rhizopus microsporus DMKU 33 in shake-flasks at 40 °C. The fermentation was then studied in a stirred-tank bioreactor with aeration at 0.75 vvm and agitation at 200 rpm, achieving the highest lactic acid production of 84 g/L with a yield of 0.84 g/g at pH 5.5 in 3 days. Lactic acid production was further increased to 105–118 g/L with a yield of 0.93 g/g and productivity of 1.25 g/L/h in fed-batch fermentation. R. microsporus DMKU 33 is thus advantageous to use in simultaneous saccharification and fermentation for l-lactic acid production from low-cost starchy substrates.     

Physiological and technological aspects of large-scale heterologous-protein production with yeasts

Commercial production of heterologous proteins by yeasts has gained considerable interest. Expression systems have been developed forSaccharomyces cerevisiae and a number of other yeasts. Generally, much attention is paid to the molecular aspects of heterologous-gene expression. The success of this approach is indicated by the high expression levels that have been obtained in shake-flask cultures. For large-scale production however, possibilities and restrictions related to host-strain physiology and fermentation technology also have to be considered. In this review, these physiological and technological aspects have been evaluated with the aid of numerical simulations. Factors that affect the choice of a carbon substrate for large-scale production involve price, purity and solubility. Since oxygen demand and heat production (which are closely linked) limit the attainable growth rate in large-scale processes, the biomass yield on oxygen is also a key parameter. Large-scale processes impose restrictions on the expression system. Many promoter systems that work well in small-scale systems cannot be implemented in industrial environments. Furthermore, large-scale fed-batch fermentations involve a substantial number of generations. Therefore, even low expression-cassette instability has a profound effect on the overall productivity of the system. Multicopy-integration systems may provide highly stable expression systems for industrial processes. Large-scale fed-batch processes are typically performed at a low growth rate. Therefore, effects of a low growth rate on the physiology and product formation rates of yeasts are of key importance. Due to the low growth rates in the industrial process, a substantial part of the substrate carbon is expended to meet maintenance-energy requirements. Factors that reduce maintenance-energy requirements will therefore have a positive effect on product yield. The relationship between specific growth rate and specific product formation rate (kg product·[kg biomass]^–1·h^–1) is the main factor influencing production levels in large-scale production processes. Expression systems characterized by a high specific rate of product formation at low specific growth rates are highly favourable for large-scale heterologous-protein production.     

Functional properties of pregelatinized and cross-linked cassava starch obtained by extrusion with sodium trimetaphosphate

Cassava starch was cross-linked with sodium trimetaphosphate (STMP) on a Cerealtec single-screw extruder at different extrusion temperatures and concentrations of STMP and NaOH. The effect of variables on functional properties of the products was analysed by the response–surface methodology. The degree of substitution (DS) was influenced by NaOH and phosphorus level, and increased when their concentration increased. Extrusion temperature affected water absorption, cold viscosity and gel characteristics. The introduction of phosphate groups by cross-linking, with maximum DS of 1.5×10⁻⁴, increased the gel strength, water absorption index, resistance to high temperature and shear, and decreased gel cohesiveness, starch clarity and water solubility index. The products had different DS and degree of gelatinization and thus can be applied in several kinds of foods.     

A semimechanistic mathematical model for growth of Rhizopus oligosporus in a model solid-state fermentation system

A Semimechanistic mathematical model is developed which describes the growth of Rhizopus oligosporus in a model solid-state fermentation system. Equations are presented for the release of glucoamylase, the diffusion of glucoamylase, the hydrolysis of starch, the generation and diffusion of glucose, and the uptake of glucose and conversion into new biomass. Good agreement of the model with the experimental data was obtained only after the glucoamylase diffusivity and the maximum specific glucose uptake rate were altered from their originally determined values. The model recognizes the distributed nature of the solid-state fermentation and therefore is able to predict the concentration profiles of the system components within the substrate. The model provides an insight into the possible rate-limiting steps in solid-state fermentation-the generation of glucose within the substrate and the resulting availability of glucose at the surface.     

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%。     

Technological aspects of extractive fermentation using aqueous two-phase systems

World Journal of Microbiology and Biotechnology -     

麦白霉素发酵培养基的改进

研究了淀粉水解液全部替代葡萄糖作为碳源发酵麦白霉素的工艺条件。用正交实验得出摇瓶培养基消毒后的最佳基质浓度为总糖5.0g/100mL,还原糖3.8g/100mL,氨基氮60mg/mL,溶解磷300μg/mL。以50L发酵罐进行实验,得出了麦白霉素的发酵代谢曲线和淀粉的适宜水解条件。经30t发酵罐放大实验结果表明,与葡萄糖作碳源相比,用淀粉水解液作碳源发酵麦白霉素,发酵单位提高23.0％,生产成本降低38.0％。     

Microbiology of the malolactic fermentation: Molecular aspects

Abstract Malolactic fermentation conducted by lactic acid bacteria follows alcoholic fermentation during winemaking, and several positive effects make it indispensable for most wines. Research has focused on the growth and physiology of lactic acid bacteria in wine; resulting in the design of malolactic starter cultures. Future work on these starters will concentrate on aromatic changes as additional criteria for strain selection. Although the main features of the malolactic enzyme and its gene are known, the detailed mechanism of the malolactic reaction remains unclear. Cloning and expression of this activity in enological strains of Saccharomyces cereuisiae might be one of the next most important advances in the control of malic acid degradation in wine.     

改革流加糖工艺提高谷氨酸发酵水平的研究

采用天津短杆菌Ｔ６－１３经多年经后的３２８＾＃菌株,以淀粉糖为糖质材料,生物素亚适量,当发酵残糖约为６０ｇ／Ｌ时,流加糖,效果好。在２００ｍ＾３发酵罐生产中,流加糖浓度分别为３００ｇ／Ｌ和４５０ｇ／Ｌ发酵周期３２小时,平均产酸分别达１０２．３ｇ／Ｌ和１１５ｇ／Ｌ,糖酸转化率５５．５％,提取收率９５．８％;单罐产理论谷氨酸由１６．２４吨提高至１８．２０吨。     

Engineering aspects of solid state fermentation

Engineering aspects of solid state fermentation have not been available in spite of the recent interest in the technique and the appearance of a number of reviews on this subject. The present state of the art regarding substrate uptake, oxygen transfer, growth characteristics, growth estimation, control systems for maintaining parameters, mathematical models, design of fermenters and automation of fermentations does not provide a detailed insight. The work carried out at the Central Food Technological Research Institute, Mysore, on the development of a large-scale solid state fermenter, comparative cost estimation of SSF and submerged fermentation as well as development of know-how for production of a variety of food enzymes, has led to the commercial exploitation of the technology by industry. Future R&D needs in this area, neglected at present but yet so promising, are indicated.     

玉米原料无蒸煮酒精发酵工艺的研究

在玉米原料无蒸煮酒精发酵过程中,添加少量的纤维素酶,酸性蛋白酶可提高糖化酶对生淀粉的糖化作用,减少糖化酶用量。在料水比１：２．５,糖化酶加量２００ｕ／ｇ,纤维素酶加量５ｕ／ｇ,酸性蛋白酶加量０．０１％,３０℃,ｐＨ３．５条件下,Ｎｏ．２１４菌株经９６ｈ发酵,醪液酒精度达１２．８％,淀粉利用率达９２．１％。     

Application of bipolar electrodialysis on recovery of free lactic acid after simultaneous saccharification and fermentation of cassava starch

The efficiency of bipolar electrodialysis (BED) for the recovery of lactic acid from fermentation broth was evaluated. Three systems of BED (bipolar-anion, bipolar-cation and bipolar-anion-cation) at fixed voltage (20 V) were compared using a model solution of ammonium lactate (100 g l(-1)). Results showed that bipolar-anion (BED-anion) was the most beneficial in terms of lactate flux, current efficiency, energy consumption and recovery ratio. Consequently, BED-anion was used to purify lactic acid from fermentation broth which had been pre-treated with mono-polar electrodialysis (MED). The final lactic acid concentration and lactate flux obtained were 144 g l(-1) and 393 g m(-2) h(-1), respectively. Using the two-step process (MED and BED-anion) the concentration of fermentation broth was increased by 33% and the total energy consumption was 2.76 kW h kg(-1).     

固态发酵工程研究进展

最近十年中 ,由于能源危机与环境问题的日益严重 ,曾被人们冷落的固态发酵再次引起人们的兴趣 ,固态发酵工程在基质特性、染菌控制、水活度的控制、pH的调控、传质与传热等领域的研究取得了较大的进展。论文着重综述最近固态发酵工程在上述领域取得的一些重大的发展 ,探讨了固态发酵过程控制参数特征及其控制策略。     

Biochemical aspects of stuck and sluggish fermentation in grape must

Recently a number of studies have focused on the factors responsible for the occurrence of stuck and sluggish fermentations. Results from these studies indicate that together with nutritional deficiencies and inhibitory substances, technological practices could lead to such situations. This review explains, from a biochemical point of view, the influence of nutritional deficiencies, inhibitory substances and technological practices on yeast cell development and physiology and the fermentation process. Received 07 February 1997/ Accepted in revised form 01 July 1997     

Mechanisms of increased linamarin degradation during solid-substrate fermentation of cassava

Several fungi and bacteria, isolated from Ugandan domestic fermented cassava, released HCN from linamarin in defined growth media. In 72 h, a Bacillus sp. decreased the linamarin to 1% of initial concentrations, Mucor racemosus to 7%, Rhizopus oryzae and R. stolonifer to 30%, but Neurospora sitophila and Geotrichum candidum hardly degraded the linamarin. Adding pectolytic and cellulolytic enzymes, but not linamarase, to root pieces under aseptic conditions, led to root softening and significantly lower linamarin contents. Neurospora sitophila showed no linamarase activity, in contrast to M. racemosus and Bacillus sp., both of which were less effective in root softening and decreasing the root linamarin content. The most important contribution of microorganisms to linamarin decrease in the solid-substrate fermentation of cassava is their cell-wall-degrading activity, which enhances the contact between endogenous linamarase and linamarin.A.J.A. Essers and M.H.J. Bennik were and M.J.R. Nout is with the Department of Food Science, Wageningen Agricultural University, Bomenweg 2, 6703HD Wageningen, The Netherlands. A.J.A. Essers is now with the Department of Toxicology, Wageningen Agricultural University, PO Box 8000, 6700EA Wageningen, The Netherlands; M.H.J. Bennik is now with the Agrotechnological Research Institute, PO Box 17, 6700AA Wageningen, The Netherlands.     

固态发酵降低上部低次烟叶中淀粉及蛋白质的含量

利用固态发酵的方法降低上部低次烟叶中淀粉和蛋白质的含量,并对发酵过程中的厌氧细菌和酵母的数量进行检测。采用单因素和正交试验对固态发酵的条件进行优化,结果表明:各因素对发酵上部低次烟叶影响显著性主次次序依次为发酵时间(C)、发酵温度(A)和发酵水分质量分数(B),固态发酵的最佳条件为A2B2C3,即温度45℃、水分质量分数50%、发酵时间9 d。在该发酵条件下,上部低次烟叶的淀粉降解率为20.41%,蛋白质降解率为12.35%。通过固态厌氧发酵的方法可取得较好的、短期内快速降解上部低次烟叶中淀粉和蛋白质含量的效果。