齐整小核菌SCL2010产小核菌多糖培养基优化的研究

采用单因素和正交试验对小核菌多糖高产菌株齐整小核菌（Sclerotium rolfsii）SCL2010的培养基成分进行了深入地筛选与优化。结果表明,最佳培养基配方为葡萄糖45.0 g/L,玉米浆1.5 g/L,NaNO3 2.0 g/L,K2HPO4·3H2O 2.0 g/L,柠檬酸0.7 g/L,MgSO4·7H2O 1.5 g/L,KCl 2.0 g/L。在此优化条件下,小核菌多糖的产量为31.81 g/L,碳源转化率为70.69%。采用发酵罐进行小试放大试验,小核菌多糖的产量达到31.86 g/L,碳源转化率为70.80%,发酵液表观黏度达到4 500 mPa·s,并将发酵时间缩短至60 h左右,具有显著效果。     

Production of carotenoids by Rhodotorula glutinis MT-5 in submerged fermentation using the extract from waste loquat kernels as substrate

BACKGROUND: The aim of present study was to investigate the feasibility of the hydrolysate extracts from waste loquat kernels as substrate in submerged culture of yeast Rhodotorula glutinis MT‐5 for carotenoid production. RESULTS: Loquat kernel was found to have high protein (22.5%) and total carbohydrate (71.2%) contents. Dried and powdered loquat kernels were subjected to acid hydrolysis with 2 mol L^?1 HCl. The hydrolysate obtained was used for the preparation of loquat kernel extract and detoxified loquat kernel extract. The detoxification of hydrolysate was performed with Ca(OH)₂. Among the 10 R. glutinis isolates, the MT‐5 was found to be best in order to produce carotenoid using the extract as substrate. Production media prepared with detoxified loquat kernel extract or loquat kernel extract gave maximum biomass concentrations of 12.64 and 11.37 g L^?1, and maximum carotenoid concentrations of 72.36 and 62.73 mg L^?1, respectively. CONCLUSION: This study has provided effective processes for the conversion of waste material of plant origin to the extracts which are very rich in term of total fermentable sugar. The practicability of the extracts as fermentation substrate was proven in carotenoid production. To the best of our knowledge, this is the first report on use of this waste material as a substrate in yeast fermentations. Copyright © 2011 Society of Chemical Industry     

Utilization of swine wastewater as a feedstock for the production of polyhydroxyalkanoates by Azotobacter vinelandii UWD

Azotobacter vinelandii UWD produced 0.69 g.l(-1) poly(hydroxybutyrate-co-hydroxyvalerate, PHBV) with 7.9 mol% hydroxyvalerate (HV) from twofold-diluted swine wastewater (SW). When supplemented with 20 g.l(-1) glucose, twofold-diluted SW increased copolymer production by 8.6 times. When three organic acids (acetate, propionate and butyrate) present in SW were supplemented with 20 g.l(-1) glucose, PHBV production was comparable (5.5 g.l(-1)) to that in the case of using twofold-diluted SW supplemented with 20 g.l(-1) glucose. However, the HV level (1.1-1.3 mol%) was very low. The component in SW contributing to copolymer production was found to be valerate. By 20 mM valerate 0.2 g.l(-1) PHBV with 44.6 mol% HV was produced. With 30 g.l(-1) glucose supplementation, 4.0 g.l(-1) PHBV with 22 mol% HV was produced. The optimal ratios of carbon to phosphorus (C : P) and to nitrogen (C : N) were 165 : 1 and 22 : 1, respectively.     

Investigation of the utility of pineapple juice and pineapple waste material as low-cost substrate for ethanol fermentation by Zymomonas mobilis

The utility of the juice of rotten or discarded pineapples and the waste material of the production of pineapple juice as low-cost substrates for ethanol production by Zymomonas mobilis was investigated. Z. mobilis ATCC 10988 produced 59.0 g.l(-1) ethanol in undiluted pineapple juice without nutritional supplementation and without the regulation of the pH while 42.5 g.l(-1) ethanol was obtained using a 125 g.l(-1) sucrose medium supplemented with 10 g.l(-1) yeast extract and mineral salts. Ethanol fermentation using unhydrolyzed and enzymatically hydrolyzed pineapple waste material was also investigated under various culture conditions. When a 15% (v/v) dilution of unhydrolyzed waste material without nutritional supplementation was used, more than 3.5 g.l(-1) ethanol was produced. When the media containing 15, 30, and 40% (v/v) of the hydrolyzate consisting of a 60% (v/v) suspension of pineapple waste material were used, final concentrations of ethanol were 5.0 g.l(-1), 7.6 g.l(-1), and 9.3 g.l(-1), respectively. These results suggest that pineapple juice and the waste material can be useful low-cost substrates for ethanol production by Z. mobilis without supplementation with expensive organic nitrogen complexes such as yeast extract and without the regulation of the pH during cultivation, leading to the reduction in the production costs.     

Identification of α‐glucosidase inhibitors from a new fermented tea obtained by tea‐rolling processing of loquat (Eriobotrya japonica) and green tea leaves

BACKGROUND: A new fermented tea produced by tea‐rolling processing of loquat (Eriobotrya japonica) leaf with green tea leaf (denoted as LG tea) showed a potent antihyperglycaemic effect in maltose‐loaded rats. The aim of this study, therefore, was to identify α‐glucosidase inhibitors in the antihyperglycaemic tea product. RESULTS: LG tea had a threefold higher maltase‐inhibitory activity (IC₅₀ 0.065 mg dried extract mL^?1) than either the constituent loquat leaf or green tea alone. In addition, LG tea favourably inhibited maltase action rather than sucrase action. As a result of bio‐guided high‐performance liquid chromatography separations of LG tea, theasinensin A, theasinensin B, strictinin and 1,6‐digalloylglucose were newly identified as maltase inhibitors with IC₅₀ values of 142, 225, 398 and 337 µmol L^?1 respectively, along with previously identified catechins and theaflavins. CONCLUSION: Judging from the magnitude of the α‐glucosidase‐inhibitory contribution of each isolated compound to the overall inhibition of LG tea, catechins were the main candidates responsible for α‐glucosidase or maltase inhibition in LG tea, followed by theaflavins, theasinensins, strictinin and 1,6‐digalloylglucose. Copyright © 2010 Society of Chemical Industry     

Suppression of blood glucose level by a new fermented tea obtained by tea‐rolling processing of loquat (Eriobotrya japonica) and green tea leaves in disaccharide‐loaded Sprague‐Dawley rats

BACKGROUND: In the field of food science, much interest has been focused on the development of alternative medicinal foods with the ability to regulate excess blood glucose level (BGL) rise. The authors have successfully developed a new fermented tea product (LG tea) by co‐fermentation of loquat (Eriobotrya japonica) leaf and summer‐harvested green tea leaf. The objective of this study was to examine the acute suppression effect of LG tea on BGL rise in disaccharide‐loaded Sprague‐Dawley (SD) rats and to evaluate its possible usage as an antidiabetic functional food material. RESULTS: As a result of single oral administration of hot water extract of LG tea (50 mg kg^?1) to maltose‐loaded SD rats, BGL at 30 min was significantly decreased by 23.8% (P < 0.01) compared with the control. A corresponding reduction in serum insulin secretion was also observed. The ED₅₀ value of LG tea (50.7 mg kg^?1) was estimated to be about 16‐fold higher than that of the therapeutic drug acarbose (3.1 mg kg^?1). CONCLUSION: No significant change in BGL was observed when sucrose or glucose was administered, suggesting that the suppression effect of LG tea was achieved by maltase inhibition, not by sucrase inhibition or glucose transport inhibition at the intestinal membrane. Copyright © 2010 Society of Chemical Industry     

利用有活力和无活力的啤酒废酵母生产酵母抽提物

以两种形态的啤酒废酵母(有活力的和无活力的)为原料,制备酵母抽提物。实验表明,采用自溶法处理啤酒废酵母液(有活力的),最佳作用条件是在温度45℃、pH 5.5下,自溶28 h,所得酵母抽提液的氨基氮含量为5.16 g/L,氨基氮得率为6.77%,产品得率为69.01%;酶采用酶水解法添加木瓜蛋白处理啤酒废酵母粉(无活力的),最佳作用条件是在木瓜蛋白酶添加量2.5%(以酵母干重计)、温度55℃p、H 4.5下,酶水解时间18 h,所得酵母抽提液的氨基氮含量为3.35 g/L,氨基氮得率为4.31%,产品得率为76.66%。     

花生粕固态发酵产纳豆激酶的工艺优化

探索纳豆菌固态发酵花生粕制备纳豆激酶的最佳工艺条件。以纳豆激酶活力为主要评价指标,在单因素实验基础上,通过正交实验优化花生粕固态发酵制备纳豆激酶的工艺条件。得出最佳发酵条件:发酵温度37℃,发酵时间24 h,接种量10%,料液比1∶0.4 g/m L。在此条件下测得发酵产物的纳豆激酶活力达到3162 U/m L,比单因素最高酶活提高了18%(2680 U/m L),可溶性氮含量为5.6 mg/m L,羟自由基清除率为74%,铁还原力的OD值为0.906。      

Utilization of shrimp shellfish waste as a substrate for solid-state cultivation of Aspergillus sp. S1-13: evaluation of a culture based on chitinase formation which is necessary for chitin-assimilation

The utilization of shrimp shellfish waste as a substrate for solid-state cultivation of a filamentous fungus, Aspergillus sp. S1-13, was investigated. The organism was selected from among 220 isolates based on the productivity of its chitinolytic enzyme (chitinase), which might reflect microbial growth. The enzyme was produced only when the organism was grown on medium containing the shellfish waste. The addition of 58-65% water (w/w) to the medium was effective in enhancing production, and a certain amount of enzyme was observed in media of higher water content (up to about 75%). The initial pH and nitrogen source (ammonium sulfate) of the solid-state medium also affected the amount of enzyme. The amount of enzyme increased 2-fold in an optimum solid-state medium: 5 g of shrimp shellfish waste and 3 ml of basal medium (pH 5) containing 0.1% (NH4)2SO4 was inoculated with 4 ml of spore suspension; static cultivation at room temperature. The amount increased further (1.5-fold) when the cultivation was carried out at 37 degrees C, with 1.85 units of the enzyme formed from 1 g of shrimp shellfish waste. An analysis by ion-exchange column chromatography suggested the presence of at least two colloidal chitin-hydrolyzing enzymes and one p-nitrophenyl beta-D-N-acetylglucosaminide-hydrolyzing enzyme in an extract of the solid-state culture. The elution profile was similar to that obtained with a liquid culture filtrate.     

Production of bacteriocin‐like inhibitory substances (BLIS) by Bifidobacterium lactis using whey as a substrate

The objective of this work was to evaluate the production of bacteriocin‐like inhibitory substances (BLIS) by Bifidobacterium animalis subsp. lactis in whey supplemented with yeast extract, inulin, Tween‐80 or l ‐cysteine. Cell growth, acidification, glucose and lactose consumption as well as BLIS production were measured during fermentations carried out in shake flasks. The best additive for both cell growth and BLIS production was shown to be yeast extract, which gave the highest concentrations of biomass (9.9 log cfu/mL) and BLIS (800 AU/mL). In a bench‐scale fermentor, B. lactis growth and BLIS production were between 6% and 25% higher than in flasks depending on the conditions assayed.     

Identification of (−)-epicatechin as the direct substrate for polyphenol oxidase from longan fruit pericarp

Postharvest browning of longan fruit results in a short life and a reduced commercial value. The experiments were conducted to separate, then purify and finally identify the polyphenol oxidase (PPO) substrates that cause longan fruit to brown. PPO and its substrates were, respectively, extracted from longan fruit pericarp tissues. The substrate for longan PPO was separated and purified using polyamide column chromatography, Sephadex LH-20 column chromatography and silica gel column chromatography, respectively. The substrate was further identified by 0.5% FeCl₃ solution and enzymatic reaction with longan PPO. On the bases of UV, ¹H NMR, ¹³C NMR, and ESI-MS data, the direct substrate for the PPO from pericarp tissues of longan fruit was identified to be (−)-epicatechin. Furthermore, the contents of (−)-epicatechin of pericarp tissues of longan fruit of two major cultivars were determined by high performance liquid chromatography (HPLC). The HPLC analysis exhibited that the contents of (−)-epicatechin of fruit pericarp of ‘Shixia’ and ‘Chuliang’ were 0.26 and 0.56 mg/g on fresh weight (FW) basis at harvest and 0.15 and 0.09 mg/g FW after 3 days of storage. The more rapid decrease in the (−)-epicatechin content of ‘Chuliang’ was due to the oxidization catalyzed by PPO, which was in agreement with the higher browning index.     

Utilization of oxidative pressure for enhanced production of poly-beta-hydroxybutyrate and poly(3-hydroxybutyrate-3-hydroxyvalerate) in Ralstonia eutropha

Ralstonia eutropha was cultivated under oxidative conditions in the presence of hydrogen peroxide and methyl viologen to stimulate the flux of NADPH to the PHA biosynthesis pathway. The effects of oxidants on the biosynthesis of poly-beta-hydroxybutyrate(PHB) and poly(3-hydroxybutyrate-3-hydroxyvalerate)[P(3HB-3HV)] were investigated. The biosynthesis rate and concentrations of PHB and its copolymer P(3HB-3HV) increased significantly under the oxidative pressure of methyl viologen, meanwhile, the molar fraction of 3HV in P(3HB-3HV) remained constant. The effect of methyl viologen on the flux of the intermediate compounds was investigated by measuring the activity of enzymes related to PHB biosynthesis, glucose 6-phosphate dehydrogenase (G6PD), and NADPH level. The oxidant significantly activated the G6PD of R. eutropha, increasing the NADPH level used for the reduction reaction of acetoacetyl-CoA to beta-hydroxybutyryl-CoA, thereby enhancing the biosynthesis of PHB and P(3HB-3HV).     

A systematic approach for extraction of phenolic compounds using parsley (Petroselinum crispum) flakes as a model substrate

The impact of extraction methodology and polarity of extraction solvents on the assay of phenolic compounds was investigated using parsley (Petroselinum crispum) flakes as a model substrate. This systematic study was undertaken to address substantial variations in the extraction procedures, solvents and conditions as described in the recent literature. Five different extraction procedures [shaking, vortex mixing, sonication, stirring and pressurized liquid extraction (PLE)] and three different solvents (methanol, ethanol and acetone), with five different solvent to water ratios per solvent, were used for extraction. Extracts were analyzed for phenolic content by high‐performance liquid chromatography and Folin–Ciocalteu assays. The yields of phenolic compounds extracted with a pressurized liquid extractor were comparable to or better than those of four classical extraction procedures. Optimum extraction efficiency with PLE was obtained when extractions were performed with four extraction cycles using ethanol–water (50:50, v/v). The amount of apiin (4,5,7‐trihydroxyflavone 7‐apiosylglucoside) and malonylapiin (apigenin malonylapiosylglucoside) isolated from parsley varied with the composition of extraction solvent. Apiin extractability was found to be a maximum when the solvent (ethanol, methanol or acetone) to water ratio was 30:70 (v/v), whereas higher amounts of malonylapiin were isolated with a reverse solvent to water ratio (70:30, v/v). Malonylapiin was not detected when parsley samples were extracted with organic solvent to water ratios of 10:90 (v/v) and 30:70 (v/v). Published in 2006 by John Wiley & Sons, Ltd     

The cultured rodent follicle as a model for investigations of gonadotrophin surge-attenuating factor (GnSAF) production

Gonadotrophin surge-attenuating factor (GnSAF) bioactivity (the suppression of GnRH-induced but not basal LH and FSH secretion from pituitary gonadotrophs) is produced by granulosa cells in vitro. Previous studies to investigate this bioactivity used dispersed granulosa cells which lack some cell types and the structural components of the follicle in vivo. The aim of this study, therefore, was to investigate whether intact rodent follicle culture was a suitable model for the study of the production of GnSAF bioactivity, allowing GnSAF to be investigated in a more physiologically realistic environment while still retaining culture conditions from which, as with granulosa cell cultures, extraneous factors can be excluded. Follicles from 16-day-old rats and 21-day-old mice were cultured for 3-6 days in the presence or absence of FSH and/or LH. The follicle-conditioned medium, and matching samples of unconditioned culture medium were added to our established rat pituitary monolayer GnSAF bioassay. Both mouse and rat intact follicles produced GnSAF bioactivity, reducing GnRH-induced LH secretion significantly. GnSAF output from the mouse follicles was highest during days 1-3 of culture, when follicles were at an early antral stage of development, and fell on days 4-6 as the follicles grew to the mid antral stage. While the stimulatory effects of FSH on rat follicle GnSAF secretion was dose-dependent, LH alone did not increase GnSAF production. An antibody against human GnSAF blocked GnSAF bioactivity produced by rat follicles, and recognised proteins within the expected pI and molecular weight range for GnSAF in two-dimensional gels of rat follicle-conditioned medium, showing a good homology between rodent and human GnSAF proteins. In conclusion, the release of GnSAF bioactivity is principally from small follicles stimulated by FSH. Therefore, intact rodent follicle culture systems offer an excellent model for the investigation of factors controlling GnSAF production under relatively physiological conditions.     

高压蒸汽提取啤酒废酵母还原型谷胱甘肽的工艺优化

为了得到高压蒸汽提取废酵母中还原型谷胱甘肽的最佳工艺条件,利用Box-Behnken的中心组合设计及响应面法（RSM）探讨了表压、提取时间、液料比和提取次数四因素的优化组合。通过建立二次回归模型,确定其最佳提取工艺条件为:表压1.1MPa,提取时间15min,液料比10∶1,提取次数1次,在此条件下得率最大为4.63mg/g。结果表明高压蒸汽提取技术是提高啤酒废酵母还原型谷胱甘肽得率的有效途径之一。      

Use of perfused isolated muscle, as studied by (31)P NMR, to investigate metabolism and post-mortem changes

An experimental system was designed to study as independently as possible the effects of various in-vivo or post-mortem factors susceptible to influence muscle metabolism. This system was made up of an NMR probe, a physiological stimulator, a perfusion system and a force monitoring device. Rabbit muscles were isolated and perfused with bovine red cells, then put into the NMR probe to follow the evolution of pH and phosphorylated compounds. It was possible to keep muscle metabolism stable for 2 h. Death was simulated by stopping the perfusion which allowed post-mortem changes to be followed. The effects of adrenaline perfusion or of a 5 s tetanus on some traits of metabolism and on changes following muscle death were studied. Tetanus immediately before perfusion was stopped accelerated changes in pH and in phosphocreatine and ATP contents; adrenaline perfusion during 30 min before perfusion was stopped had little effect on these traits.