Study of operational variables in the submerged growth ofPleurotus ostreatus mushroom mycelium

Pleurotus ostreatus mushroom mycelium was cultivated in submerged culture in shake-flask experiments with acid extract from peat and yeast extract as nutrient sources. Different concentrations of water-diluted peat extract were tested in an attempt to overcome the effect of growth inhibitors apparently present in nondiluted peat extracts. The best results were obtained with a ratio of one part of peat extract diluted with one part of water. Several operating variables were studied to optimize the growth of mycelial biomass ofP. ostreatus. The best results produced approximately 5 g/L dry biomass with a yield of 60% and an efficiency of 33%. These results were obtained in 8 d at 5% (v/v) inoculum ratio, 28°C, pH of 5.0, and 150 rpm.     

HPLC quantification of rDNA polypeptides like insulin precursors produced in yeast

Summary A reversed-phase HPLC system for the quantification of recombinant DNA insulin precursors in yeast fermentation broth is described. HPLC monitoring of the polypeptide can be achieved without interference from substrate components or proteins stemming from the host cell, thus serving to determine the optimum harvesting time. Furthermore, it is demonstrated how different amino acid links of the insulin A- and B-chains affect the chromatographic behaviour of the insulin precursor making it possible to identify each precursor by its retention on the reversed phase support.     

SnO2的制备及催化臭氧氧化活性

采用沉淀法制备了SnO2催化剂,以SnO2催化臭氧氧化降解糖蜜酒精废水脱色为探针反应,对催化剂的活性进行了评价.采用X射线衍射、红外光谱及热分析(TG-DSC)等技术对催化剂进行表征,研究了沉淀剂及焙烧温度等制备参数对SnO2催化臭氧氧化活性的影响.结果表明,SnO2催化剂对臭氧氧化降解糖蜜酒精废水脱色具有较高的催化活性,反应60 min后,糖蜜酒精废水的脱色率从单独臭氧氧化的43.04%提高到60.24%.沉淀剂对SnO2催化剂的活性影响很大,其中以氨水为沉淀剂制备的SnO2催化剂去羟基化反应程度高,所制得的催化剂活性最大.催化剂适宜的焙烧温度为723 K.SnO2吸附吡啶的红外光谱表明,催化剂表面存在L酸中心.臭氧在SnO2表面吸附的红外光谱表明,通过臭氧的末端氧原子与表面羟基及L酸中心成键,生成的活性氧可氧化降解糖蜜酒精废水.     

Potential Application of Alkaline Pectinase from <Emphasis Type="Italic">Bacillus subtilis</Emphasis> SS in Pulp and Paper Industry

Pectinase production from Bacillus subtilis SS was optimized under solid-state fermentation (5,943 U/g of dry bacterial bran). The pectinase produced was stable in neutral to alkaline pH range at 70 degrees C; therefore, the suitability of this pectinase in pulp and paper industry was investigated. The enzyme pretreatment process was optimized, and a pectinase dose of 5 IU/g of oven-dried pulp (10% consistency) at pH 9.5 temperature 70 degrees C after 150 min of treatment gave the best pretreatment to the pulp. An increase of 4.3% in brightness along with an increase of 14.8 and 65.3% in whiteness and fluorescence, respectively, whereas a 15% decrease in the yellowness of the pretreated pulp were observed. There was a 5.85% reduction in kappa number and 6.1% reduction in permanganate number along with a reduction in the chemical oxygen demand value. Significant characteristics showed by pectinase open new possibilities of application of this cellulase-free enzyme in the pulp and paper industry by reducing the negative environmental impact of chemicals apart from improving the properties of paper.     

A Comparison of Simple Rheological Parameters and Simulation Data for Zymomonas mobilis Fermentation Broths with High Substrate Loading in a 3-L Bioreactor

Traditionally, as much as 80% or more of an ethanol fermentation broth is water that must be removed. This mixture is not only costly to separate but also produces a large aqueous stream that must then be disposed of or recycled. Integrative approaches to water reduction include increasing the biomass concentration during fermentation. In this paper, experimental results are presented for the rheological behavior of high-solids enzymatic cellulose hydrolysis and ethanol fermentation for biomass conversion using Solka Floc as the model feedstock. The experimental determination of the viscosity, shear stress, and shear rate relationships of the 10 to 20% slurry concentrations with constant enzyme concentrations are performed with a variable speed rotational viscometer (2.0 to 200 rpm) at 40 °C. The viscosities of enzymatic suspension observed were in range of 0.0418 to 0.0144, 0.233 to 0.0348, and 0.292 to 0.0447 Pa s for shear rates up to 100 reciprocal seconds at 10, 15, and 20% initial solids (w/v), respectively. Computational fluid dynamics analysis of bioreactor mixing demonstrates the change in bioreactor mixing with increasing biomass concentration. The portion-loading method is shown to be effective for processing high-solids slurries.     

Aqueous Ammonia Soaking of Switchgrass Followed by Simultaneous Saccharification and Fermentation

Simultaneous saccharification and fermentation (SSF) of switchgrass was performed following aqueous ammonia pretreatment. Switchgrass was soaked in aqueous ammonium hydroxide (30%) with different liquid–solid ratios (5 and 10 ml/g) for either 5 or 10 days. The pretreatment was carried out at atmospheric conditions without agitation. A 40–50% delignification (Klason lignin basis) was achieved, whereas cellulose content remained unchanged and hemicellulose content decreased by approximately 50%. The Sacccharomyces cerevisiae (D₅A)-mediated SSF of ammonia-treated switchgrass was investigated at two glucan loadings (3 and 6%) and three enzyme loadings (26, 38.5, and 77 FPU/g cellulose), using Spezyme CP. The percentage of maximum theoretical ethanol yield achieved was 72. Liquid–solid ratio and steeping time affected lignin removal slightly, but did not cause a significant change in overall ethanol conversion yields at sufficiently high enzyme loadings. These results suggest that ammonia steeping may be an effective method of pretreatment for lignocellulosic feedstocks.     

Lipase Production in Solid-State Fermentation Monitoring Biomass Growth of Aspergillus niger Using Digital Image Processing

The aim of this study was to monitor the biomass growth of Aspergillus niger in solid-state fermentation (SSF) for lipase production using digital image processing technique. The strain A. niger 11T53A14 was cultivated in SSF using wheat bran as support, which was enriched with 0.91% (m/v) of ammonium sulfate. The addition of several vegetable oils (castor, soybean, olive, corn, and palm oils) was investigated to enhance lipase production. The maximum lipase activity was obtained using 2% (m/m) castor oil. In these conditions, the growth was evaluated each 24 h for 5 days by the glycosamine content analysis and digital image processing. Lipase activity was also determined. The results indicated that the digital image process technique can be used to monitor biomass growth in a SSF process and to correlate biomass growth and enzyme activity. In addition, the immobilized esterification lipase activity was determined for the butyl oleate synthesis, with and without 50% v/v hexane, resulting in 650 and 120 U/g, respectively. The enzyme was also used for transesterification of soybean oil and ethanol with maximum yield of 2.4%, after 30 min of reaction.     

Pretreatment of Whole-Crop Harvested, Ensiled Maize for Ethanol Production

To have all-year-round available feedstock, whole-crop maize is harvested premature, when it still contains enough moisture for the anaerobic ensiling process. Silage preparation is a well-known procedure for preserving plant material. At first, this method was applied to obtain high-quality animal feed. However, it was found that such ensiled crops are very suitable for bioenergy production. Maize silage, which consists of hardly degradable lignocellulosic material, hemicellulosic material, and starch, was evaluated for its potential as a feedstock in the production of bioethanol. It was pretreated at low severity (185 °C, 15 min) giving very high glucan (∼100%) and hemicellulose recoveries (<80%)—as well as very high ethanol yield in simultaneous saccharification and fermentation experiments (98% of the theoretical production based on available glucan in the medium). The theoretical ethanol production of maize silage pretreated at 185 °C for 15 min without oxygen or catalyst was 392 kg ethanol per ton of dry maize silage.     

海洋烟曲霉菌产脱乙酰酶发酵条件优化

几丁质酶是一类复合酶,包括脱乙酰酶、几丁质酶、壳聚糖酶、几丁寡糖酶、几丁二糖酶等,将脱乙酰酶应用于壳聚糖的制备中,不仅可以有效地从甲壳素(几丁质)分子中脱除乙酰基,而且不会把甲壳素的长链降解为小分子,可以制备出高脱乙酰度且性能独特的壳聚糖,同时不存在排放废碱液而对环境造成严重污染.     

A statistical approach using L<Subscript>25</Subscript> orthogonal array method to study fermentative production of clavulanic acid by <Emphasis Type="Italic">Streptomyces clavuligerus</Emphasis> MTCC 1142

Clavulanic acid is a naturally occurring antibiotic produced by Streptomyces clavuligerus. The present work reports on clavulanic acid production by Streptomyces clavuligerus MTCC 1142 using one-factor-at-a-time and L₂₅ orthogonal array. The one-factor-at-a-time method was adopted to investigate the effect of media components (i.e., carbon source, nitrogen source and inoculum concentration) and environmental factors such as pH for clavulanic acid production. Production of clavulanic acid by Streptomyces clavuligerus was investigated using seven different carbon sources (viz. glucose, sucrose, modified starch, rice-bran oil, soybean oil, palm oil, and glycerol) and six different nitrogen sources (viz. peptone, yeast extract, ammonium chloride, ammonium carbonate, sodium nitrate and potassium nitrate). A maximum yield of 140 μg/mL clavulanic acid was obtained in the medium containing soybean oil as a carbon source and yeast extract as nitrogen source. Subsequently, the concentration of soybean flour, soybean oil, dextrin, yeast extract and K₂HPO₄ were optimized using L₂₅ orthogonal array method. The final optimized medium produced 500 μg/mL clavulanic acid at the end of 96 h as compared to 140 μg/mL before optimization. Synthesis of precursor molecules as a metabolic driving force is of considerable importance in antibiotic synthesis. Attempts to increase the clavulanic acid synthesis by manipulating the anaplerotic flux on C₃ and C₅ precursors by supplementing the medium with arginine, ornithine, proline, valine, leucine, isoleucine, pyruvic acid and á-ketoglutarate were successful. Supplementing the optimized medium with 0.1 M arginine and 0.1 M leucine increased the yield of clavulanic acid further to 1100 μg/mL and 1384 μg/mL respectively.