黑曲霉新菌株HA608的诱变及选育

经诱变选育，筛选获得一株适宜于黄姜和橡子水解糖液发酵的新菌株ＨＡ６０８，糖液不需经任何处理，摇瓶发酵１１０ｈ，转化率１１０％左右。     

木质纤维素稀酸水解糖液乙醇发酵研究进展

以木质纤维素为原料生产燃料乙醇,首先要对原料进行预处理得到可发酵糖,在稀酸水解木质纤维素得到的糖液中,除含有葡萄糖、木糖等六碳糖和五碳糖外,根据水解温度、酸浓度和时间的不同,还含有不同浓度的发酵抑制剂。因此,在研究木质纤维素稀酸水解糖液的乙醇发酵中,对代谢木糖成乙醇的菌种的研究、对耐/代谢发酵抑制剂微生物的研究、对稀酸水解糖液的脱毒方法的研究以及对稀酸水解糖液不同发酵方式的乙醇发酵研究等非常重要。重点介绍了以上几个方面近几年研究的进展。     

亚硫酸盐甘蔗渣浆酶解液发酵制备乙醇

以亚硫酸盐甘蔗渣浆酶解液作为原料,利用C. shehatae发酵制取燃料乙醇。结果表明：还原糖最适初始质量浓度为葡萄糖140 g/L、木糖60 g/L、酶解液总糖80 g/L。利用初始葡萄糖55.06 g/L、木糖11.18 g/L、纤维二糖4.51 g/L的亚硫酸盐甘蔗渣浆酶解液发酵,经18 h获得乙醇22.98 g/L。乙醇得率为67.23%,葡萄糖利用率为99.27%,木糖利用率为32.96%,C. shehatae适合作为蔗渣为原料的乙醇发酵菌株。     

环境条件及摇瓶补糖策略对谷胱甘肽发酵的影响

研究了酵母摇瓶发酵中pH、装液量、初糖浓度、碳氮磷比和补糖方式对谷胱甘肽(GSH)发酵的影响。结果表明,GSH发酵适宜的初始pH和装液量分别为6 0和500ml锥形瓶内装液量60m1。初糖浓度对GSH发酵有较大的影响,超过12g／l,的初糖浓度将减少胞内GSH含量。应用计算得出的一种以控制比生长速率为目的的摇瓶补糖策略,在总糖浓度为26．2g／L下发酵12h,最终细胞浓度和胞内GSH含量分别达到8.78g／L和13．6mg／g,发酵液内GSH总量达到119．4mg／L,细胞对糖产率达到0.335g／g。     

玉米浆对玉米粉和木薯淀粉发酵甘油的影响及二者的比较

以玉米粉和木薯淀粉为原料 ,比较了二者的液化和糖化 ,结果表明 :在相同条件下 ,木薯淀粉液化时间较短 ,玉米粉液化时间较长 ,但二者的液化液均较易糖化。然后分别以玉米粉和木薯淀粉糖化液为原料 ,用耐高渗酵母发酵生产甘油 ,研究了玉米浆对二者甘油发酵的影响并对二者进行了比较 ,结果表明 :当玉米粉和木薯淀粉糖化液还原糖含量分别为 2 5 % ,尿素为 0 .2 % ,pH为 4 .5时 ,用玉米粉糖化液发酵甘油时可不添加玉米浆 ,甘油产量最高可达 2 % ,而用木薯淀粉糖化液发酵甘油时 ,适宜的玉米浆为 0 .15 % ,甘油产量最高可达 4 .9%。对二者的比较结果表明 :用玉米粉糖化液为发酵原料时 ,发酵时间较短 ,残糖降低较快 ,甘油产量较低 ,在 36h之后 ,甘油开始反耗。而用木薯淀粉糖化液发酵时 ,发酵时间较长 ,残糖降低较慢 ,甘油产量较高 ,在 72h之后 ,甘油开始反耗。     

槐米药渣固体发酵法生产畜禽饲料工艺的研究

以发酵酶解液中总糖、芦丁、槲皮素含量为实验分析指标来确定槐米残渣固体发酵生产畜禽饲料生产工艺。实验结果表明:黑曲霉槐米残渣固体发酵的最佳工艺条件为接种量4%,发酵酶解p H 5.5,温度35℃,时间5 d。槐米残渣经发酵酶解的总糖量高于未经发酵酶解的总糖量60%以上,槲皮素含量高于65%以上。槐米残渣经黑曲霉发酵酶解可作为抗菌、消炎畜牧营养饲料。     

糖液DE值对柠檬酸发酵的影响研究

本文通过调节加酶量等方法来得到不同DE值梯度的液化清液,然后进行柠檬酸发酵摇瓶试验,旨在验证糖液DE值对柠檬酸发酵的影响。     

沙柳原料高浓度底物酶解发酵乙醇工艺

为了提高沙柳生物转化过程的经济可行性,考察了沙柳原料经过蒸爆、超微粉碎+稀酸、超微粉碎+稀碱预处理后高浓度底物补料酶解的效果,并对其高浓度水解糖液进行了乙醇发酵。结果表明:蒸爆处理法水解效果最好,通过补料酶解,底物质量分数可以达到30%,酶解液中总糖质量浓度达到132 g/L,葡萄糖质量浓度105 g/L;超微粉碎+稀酸预处理原料底物质量分数可以达到22%,酶解液中总糖质量浓度达到123 g/L,葡萄糖质量浓度73 g/L;超微粉碎+稀碱预处理原料底物质量分数可以达到22%,酶解液中总糖质量浓度133 g/L,葡萄糖质量浓度77 g/L。3种预处理使沙柳原料的酶解糖液都可以较好地被酿酒酵母利用发酵产乙醇,蒸爆处理原料的酶解糖液乙醇发酵效果最好,乙醇质量浓度达到47 g/L。     

发酵工艺条件的多目标优化

以模糊综合评价值作为评价指标、加权单纯形方法优化发酵过程工艺条件。应用本方法,对以谷氨酸发酵废液为原料的酵母发酵过程的发酵温度、ｐＨ、菌种量、初始糖浓度、通气量及流加液糖浓度等工艺条件进行了优化。     

产β—甘露聚糖酶地衣芽孢杆菌的分离筛选及发酵条件

从土样中分离筛选出产β－甘露降糖酶的地衣芽孢杆菌,经紫外线诱变处理后获得高酶活力ＮＫ－２７菌株,在以魔芋粉、豆饼粉为碳氮源添加无机盐的发酵培养基中,β－甘露聚糖酶活力达１１０．４９ｕ／ｍｌ。初始ＰＨ值、装液量、培养温度和培养时间对产酶有一定影响。     

Comparative kinetics of appearance in the portal vein of alpha-aminated nitrogen from mixtures of small peptides of free amino acids of the same composition introduced in the duodenum of conscious pigs

The quantitative kinetics of appearance of amino acids (a.a.) in the pig portal vein was studied in 6 animals for 5 hrs. after duodenal perfusion of an enzymatic hydrolysate of milk proteins or a solution of free a.a. of the same composition. Each product was given in two quantities (55 and 110 g). The quantities of a.a. appearing in the portal vein were higher after perfusion of the hydrolysate than after that of the free a.a., independently of the time after the perfusion. Thus, nitrogen present in the small intestine as small peptides is absorbed more quickly than when it is present as free amino acids.     

酵母发酵蔗渣半纤维素水解物生产木糖酶

采用二次正交旋转组合设计研究了蔗渣半纤维素水解过程中硫酸浓度与液 固比对木糖收率的影响。回归分析表明 ,这两个因素与木糖的收率之间存在显著的回归关系。通过回归方程优化水解条件 ,当硫酸浓度 2 .4g L ,液 固 =6 .2 ,在蒸汽压力 2 .5× 10 4Pa的条件下水解 2 .5h ,10 0g蔗渣可水解生成木糖约 2 4g。大孔树脂吸附层析处理蔗渣半纤维素水解物 ,能有效地减少其中的酵母生长抑制物含量 ,显著改善水解物的发酵性能。用大孔树脂在pH 2条件下处理过的蔗渣半纤维素水解物作基质 ,含木糖 2 0 0g L ,产木糖醇酵母菌株CandidatropicalisAS2 .1776发酵 110h耗完基质中的木糖 ,生成木糖醇 12 7g L ,产物转化率 0 .6 4(木糖醇g 木糖g) ,产物生成速率 1.15g L·h .     

氨基酸分析仪测定饲料添加剂——叶酸的方法

用６ｍｏｌ／Ｌ盐酸于１１０℃条件下水解饲料添加剂——叶酸,使之游离出谷氨酸,用氢氧化钠中和调节ｐＨ到２,氨基酸分析仪测定谷氨酸含量,经与标准叶酸水解样品比较,计算出叶酸的纯度。该方法重现性好,变异系数ＣＶ＝０．０８％,平均回收率为９８．３４％,浓度与峰面积呈线性相关,相关系数ｒ＝０．９９８７,可随氨基酸分析同时进行,不需改变任何分析条件。     

荧光法测定羽毛中的色氨酸含量

本文探讨了羽毛中色氨酸的荧光分析方法,将２～５ｍｇ的羽毛样品置于聚四氟乙烯管中,用１ｍｌ含０．５％可溶性淀粉的５ｍｏｌ／Ｌ　ＮａＯＨ为水解液,以纯氮为保护气体,在１１０℃水解２０小时,水解产物用６ｍｏｌ／ＬＨＣｌ中和,然后用Ｎａ２ＨＰＯ４－ＮａＯＨ的缓冲溶液（ｐＨ１１）稀释５倍,在激发波长２８０ｎｍ,发射波长３６２ｎｍ测定荧光强度及其浓度。经多次试验,结果较为满意,重现性较好,其相对标准偏差为０．６１％～３．００％。样品的回收率为９５．３％～１０２．１％。此法快速、简便,适用于各种羽毛的例行分析。     

DO-stat fed-batch production of 2-keto-d-gluconic acid from cassava using immobilized Pseudomonas aeruginosa

Bioconversion of cassava-derived glucose to 2-keto-d-gluconic acid (2-KDG) using resting cells of immobilized Pseudomonas aeruginosa IFO 3448 was investigated. The tuberous roots of cassava were selected as the feedstock as they are inexpensive and widely available, and possess high amounts of starch (approximately 70% (w/w) of dry mass). Immobilized bacteria was used in a fed-batch fermenter and recycled over a period of 2 weeks. Given that the formation of 2-KDG from glucose requires oxygen as a reagent, and that high glucose concentrations are detrimental to the production yield of 2-KDG by resting cells, a DO-stat control strategy was used, whereby the feed rate of cassava hydrolysate was regulated by coupling it with the control variable, dissolved oxygen. For 319 h of operation including three cycles of repeated fed batch, 72 g of 2-KDG was produced from hydrolysate derived from 110 g of dried cassava at a maximum production rate of 0.55 g/L/h and an average concentration of 35 g/L.     

gamma-Carboxyglutamic acid in invertebrates: its identification in hermatypic corals

The presence of a γ-carboxyglutamate-containing protein in hermatypic corals has been established. γ-Carboxyglutamate has been isolated from the alkaline hydrolysate of protein extracted from the coral Lobophyllia corymbosa, by chromatography of the hydrolysate on Dowex AG 1-X8 (formate form), followed by chromatography on an amino acid analyzer column. This procedure achieves complete separation of γ-carboxyglutamate from an acid-stable compound which is also present in the alkaline hydrolysate of coral protein, and which has proved difficult to separate from γ-carboxyglutamate by other methods. The identity of the γ-carboxyglutamate thus isolated has been established unequivocally by determining the yield of glutamic acid after acid treatment (2 M HCl, 6 h, 110°C). The color factor for the γ-carboxyglutamate isolated from Lobophyllia is 0.45₄ times the value for glutamic acid, in good agreement with the value determined for authentic γ-carboxyglutamic acid (0.45₈) under exactly the same conditions. Virtually identical results have been obtained for the coral, Acropora cuneata. These experiments provide the first secure evidence for the presence of γ-carboxyglutamate in an invertebrate species, and they clearly have important implications for our understanding of invertebrate mineralization.     

瓜尔豆种皮酶解提取物抗氧化性研究

以3种蛋白酶对瓜尔豆种皮活性肽进行酶解分离,通过总抗氧化能力测定,筛选出木瓜蛋白酶水解提取物总抗氧化能力最强,分别为碱性蛋白酶和中性蛋白酶水解提取物的1．95倍和3．34倍。在清除超氧阴离子自由基的测定中,木瓜蛋白酶水解提取物也表现出较强的清除能力,清除率随水解溶液浓度的增加呈正量效关系,当溶液浓度为5．45mg/mL时,清除率达43．37％。通过实验证实瓜尔豆种皮酶解提取物与大豆多肽一样有较强的抗氧化能力。     

Enhancing effect of albumin hydrolysate on ethanol production employing Saccharomyces sake

The enhancing effect of albumin hydrolysate on ethanol production was investigated in ethanol fermentations using Saccharomyces sake. In batchwise ethanol production, addition of supplemental albumin hydrolysate and phosphatidylcholine, or albumin hydrolysate alone, brought about a more than 60% increase in final ethanol concentration (148 or 144 g/L compared with 88 g/L with no supplementation [control] after 72 h). The effect of the supplements is believed to be due to an enhanced alcohol tolerance of cells grown in media containing the supplements. Cells grown in media containing albumin hydrolysate were enriched in phenyalanine, tyrosine, and methionine in their plasma membranes. All three amino acids were also present in considerable amounts in the albumin hydrolysate. This fact suggests that the three amino acids, which are present in albumin hydrolysate, are incorporated into the plasma membranes of cells. Under ethanol production conditions in which only one amino acid among the components of albumin hydrolysate was excluded, namely phenlalanine, tyrosine, or methionine, significant reductions in ethanol production resulted. (c) 1995 John Wiley & Sons, Inc.     

Kinetic behavior of Candida guilliermondii yeast during xylitol production from Brewer's spent grain hemicellulosic hydrolysate

Brewer's spent grain, the main byproduct of breweries, was hydrolyzed with dilute sulfuric acid to produce a hemicellulosic hydrolysate (containing xylose as the main sugar). The obtained hydrolysate was used as cultivation medium by Candidaguilliermondii yeast in the raw form (containing 20 g/L xylose) and after concentration (85 g/L xylose), and the kinetic behavior of the yeast during xylitol production was evaluated in both media. Assays in semisynthetic media were also performed to compare the yeast performance in media without toxic compounds. According to the results, the kinetic behavior of the yeast cultivated in raw hydrolysate was as effective as in semisynthetic medium containing 20 g/L xylose. However, in concentrated hydrolysate medium, the xylitol production efficiency was 30.6% and 42.6% lower than in raw hydrolysate and semisynthetic medium containing 85 g/L xylose, respectively. In other words, the xylose-to-xylitol bioconversion from hydrolysate medium was strongly affected when the initial xylose concentration was increased; however, similar behavior did not occur from semisynthetic media. The lowest efficiency of xylitol production from concentrated hydrolysate can be attributed to the high concentration of toxic compounds present in this medium, resulting from the hydrolysate concentration process.