木聚糖酶Shearzyme 500 L对蔗渣木聚糖的特性研究

以木聚糖酶Shearzyme 500L水解蔗渣木聚糖制备低聚木糖,用DNS法测定酶解液中的总糖和还原糖,HPLC法测定酶解产物组成,其适宜的水解条件为底物质量浓度3g/100mL、pH5.0、60℃、木聚糖中酶用量50U/g、水解时间24h。在此条件下底物水解率约为63.1%,水解产物的81.5% 为低聚木糖,其中木二糖占54.8%,木三糖占26.7%。Shearzyme 500L 不能将一分子木二糖水解为两个木糖单糖,但能水解木三糖并相应生成木二糖与木糖。副产物木糖能显著抑制Shearzyme 500L 活性,降低木聚糖的水解率。     

Production of L-lactic acid from corncob

The optimum temperature, initial pH, amount of added enzyme and substrate (corncob) for the hydrolysis of corncob by Acremonium cellulase were 35 degrees C, 4.5, 10 u/g-corncob and 100 g/l, respectively. Under the optimum conditions, more than 55 g/l of reducing sugars were hydrolyzed from 100 g/l of corncob to 34 g/l of glucose and 12 g/l of xylose based on dried corncob. More than 25 g/l of L-lactic acid was produced from this enzymatic hydrolyzate and less than 5 g/l of xylose remained in the 3-l airlift bioreactor. The production of L-lactic acid by simultaneous saccharification and fermentation (SSF) was also carried out in the 3-l airlift bioreactor using Acremonium thermophilus (cellulose-producer) and Rhizopus sp. MK-96-1196 (lactic acid-producer). More than 24 g/l of L-lactic acid was produced from 100 g/l of untreated raw corncob.     

Interaction between Lactococcus lactis and Lactococcus raffinolactis during growth in milk: development of a new starter culture

Many milk fermentations use mixed cultures of lactic acid bacteria. To select a new mixed starter culture, 100 acid-producing bacterial strains were isolated from raw cow milk. Of these, 13 strains identified as belonging to the genera Lactococcus, Lactobacillus, Leuconostoc, or Weissella (based on phenotypic and genotypic tests) were assessed for a symbiotic effect between pairs of isolated strains during growth in milk. Among the strains tested, a mixed culture of Lactococcus lactis ssp. lactis strain 54 and Lactococcus raffinolactis strain 37 stimulated greater acid production during fermentation than occurred with pure fermentation. This stimulatory effect was not observed in milk supplemented with yeast extract or glucose or in constituted medium. Addition of a cell-free filtrate from milk fermented by strain 54 increased acid production by strain 37; however, the converse effect was not observed. The increased acid production by this mixed culture was, therefore, due to stimulation of strain 37 by metabolic products of strain 54, suggesting that the interaction between strains 54 and 37 is commensal. Analysis with a taste-sensing system indicated that fermented milk containing the mixed culture was more acidic, had more anionic bitterness, had greater aftertastes of anionic bitterness and astringency, and was less salty and umami than milk containing the individual cultures. This study identifies a new commensal relationship between 2 lactococcal strains that are commonly used for making dairy products.     

Production of lactic acid from xylose and wheat straw by Rhizopus oryzae

Rhizopus oryzae NBRC 5378 was the best among 56 strains of R. oryzae for the production of lactic acid from xylose. This strain produced lactic acid from wheat straw powder by a simultaneous saccharification and fermentation process, with a yield of 0.23 g/g from cellulose and hemicellulose in wheat straw.     

混合糖发酵产油脂酵母菌的筛选

对6株产油脂酵母利用葡萄糖、木糖及不同比例混合糖(葡萄糖与木糖)发酵的菌体生长和产油特性进行研究,筛选到一株能够转化混合糖的高产油脂酵母JM-D.该菌株在混合糖比例为3∶1的条件下发酵120h,菌体油脂含量可达23.38％,油脂产量可达2.7g/L;菌体油脂中饱和脂肪酸含量占脂肪酸总量的49.2％;混合糖比葡萄糖更容易被转化生成饱和脂肪酸含量较多的油脂.     

甘肃传统浆水发酵过程中产双戊烯乳酸菌的分离筛选

目的:从传统发酵浆水中分离筛选产双戊烯乳酸菌株,以期为传统浆水工艺现代化改造过程增香发酵剂的开发提供菌株来源。方法:采用经典的平板分离筛选方法初筛菌株,结合发酵性能测定复筛。结果:从甘肃传统发酵浆水中初筛得到5株产双戊烯乳酸菌,通过测定其发酵产双戊烯性能,从中复筛得到1株产双戊烯乳酸菌菌株R-32,该菌在马铃薯番茄液体培养基中产双戊烯含量可达到0.171 μg/mL;通过观察其形态、测定生理生化指标及进行产乳酸的定性试验,鉴定该菌株R-32为肠膜明串珠菌属(Leuconostoc mesenteroides)。结论:本实验获得了一株可用于传统浆水发酵的产香乳酸菌株。     

不同凝结芽孢杆菌在单一及混合碳源下的发酵特性

本文研究了凝结芽孢杆菌BH1、HL5和36D1在木糖、海藻糖、葡萄糖、低聚异麦芽糖、蔗糖和麦芽糖等单一碳源和葡萄糖分别加其他五种碳源(即混合碳源)下的底物利用和代谢情况,对三株菌以葡萄糖和低聚异麦芽糖作为混合碳源进行5 L罐发酵,进一步考察三株菌发酵特性,以期确定一株菌株作为后续高效乳酸发酵菌株筛选的出发菌株。结果显示:单一碳源发酵下,凝结芽孢杆菌36D1可以完全利用海藻糖、木糖和葡萄糖,而BH1和HL5对木糖和海藻糖几乎无利用,同时三株菌对蔗糖均无明显利用;混合碳源发酵下,凝结芽孢杆菌36D1发酵过程中表现出明显的葡萄糖阻遏效应,BH1和HL5对其他糖(尤其是麦芽糖和异麦芽糖)的利用几乎不受影响;5 L罐发酵结果表明,HL5对异麦芽糖的利用能力较优。凝结芽孢杆菌HL5对麦芽糖、异麦芽糖以及葡萄糖的利用能力较高,最终确定HL5作为后续菌株筛选的出发菌株。     

橄榄绿链霉菌E-86产木聚糖酶水解玉米芯汽爆液生产低聚木糖

本文以橄榄绿链霉菌E-86产木聚糖酶水解玉米芯汽爆液生产低聚木糖为目的,研究了玉米芯汽爆液的水解特征,并与玉米芯木聚糖的酶解产物组成进行了比较,得到如下结果:加酶量120U/100ml、酶解反应8h可获得较好的酶解效果,直接还原糖量46μmol/ml、平均聚合度3.1、水解率46%;玉米芯汽爆液和玉米芯木聚糖的酶解产物中低聚糖组成大致相同,主要是木二糖和少量的木三糖、木糖,汽爆液酶解产物中还含有极少量的鼠李糖和阿拉伯糖;玉米芯汽爆液可代替玉米芯木聚糖为底物生产低聚木糖。     

发酵乳球菌菌株的分离鉴定

对9株发酵乳球菌菌株进行MRS固体平板培养基划线分离和形态学比较观察，井采用生化试验和RAPD对其进行鉴定。划线分离得到11株菌：通过菌体常规磷钨酸负染色标本的透射电镜观察，描述了各菌体的大小、形状以及裂殖方式；依据各菌种生化代谢差异设计生化试验．得到了2个发酵型，将其鉴定为2个菌群（乳酸乳球菌乳酸亚种和嗜热链球菌）；20个随机引物的RAPD筛选得到了9个不同的基因型．将其鉴定为9株细菌。     

酒曲中乳酸菌的筛选及其在板栗糯米饮料发酵中的应用

为筛选出适合植物基质发酵的优良乳酸菌.本文以米曲为目标乳酸菌的供体,筛选高产酸、耐胃肠环境能力良好的乳酸菌,并评估了乳酸菌的抗氧化活性及其在板栗糯米饮料中的发酵能力.从酒曲中共分离出84株乳酸菌,其中有38株的产酸量≥10 g/L;在进一步的耐酸耐胆盐、模拟胃肠实验中,共筛出3株乳酸菌株;经16S rDNA鉴定这3株乳...     

嗜热拟青霉木糖苷酶的性质及其与木聚糖酶的协同作用

嗜热拟青霉(Paecilomyces thermophila)J18利用玉米芯能产胞外木糖苷酶,通过(NH_4)_2SO_4沉淀、DEAE-52离子交换层析及Q-琼脂糖凝肢-FF(QSFF)离子交换层析从上清液纯化得到了电泳纯的木糖苷酶,纯化倍数为31.9倍,回收率为2.27%。SDS-PAG E及Superdex-75凝胶过滤层析测定木糖苷酶的分子量分别为53.5 ku和51.8 ku。该酶最适温度及pH分别为55℃和pH 6.5。木糖苷酶能够水解木二糖和低聚木糖但不能水解木聚糖,水解低聚木糖的相对速率随聚合度增加而增加。该酶对木糖的抑制常数Ki值为139 mmol/L,具有很高的木糖耐受性。木糖苷酶与内源木聚糖酶一起水解木聚糖产生更多的还原糖,表现出协同作用。     

双菌微囊化发酵耦合泡沫分离强化乳链菌肽生产的工艺

本研究拟通过共培养乳酸链球菌与酿酒酵母解除产物抑制效应,并将泡沫分离耦合于微胶囊固定细胞发酵强化乳链菌肽（Nisin）的生产。采用液芯微胶囊固定乳酸链球菌和酿酒酵母细胞,并对海藻酸钠和壳聚糖浓度进行优化。结果表明,以乳糖为底物碳源,乳酸链球菌与酿酒酵母的初始接种比例102:1时,共培养发酵液中Nisin效价高达3436.3 IU/mL。海藻酸盐-壳聚糖-海藻酸盐（ACA）液芯微胶囊的制备条件为海藻酸钠浓度15 g/L和壳聚糖浓度5.5 g/L。在微胶囊固定细胞发酵与泡沫分离耦合时间为21 h,分布器孔径180 μm,气体体积流率40 mL/min条件下,Nisin的富集比和回收率分别为5.8和63.1%,总效价为3973.2 IU/mL。本研究建立的双菌微囊化发酵-泡沫分离耦合工艺能够有效解除产物抑制和菌体细胞损失,为高密度、连续发酵生产Nisin奠定了理论基础。     

眉山泡菜中乳酸菌的分离鉴定

目的：分析眉山泡菜的基本数据和乳酸菌的菌种构成。方法：从眉山市采集泡菜12 份,测定pH值、总酸度、盐度、乳酸菌计数,用16S rRNA序列分析法鉴定菌种。结果：样品pH值的范围是3.37～3.89,总酸度的范围是0.65～0.92 g/100 g（以乳酸计）,盐度的范围是4.16%～5.28%（以NaCl计）,MRS和M17乳酸菌计数分别是1.71～6.33、1.33～5.78（lg（CFU/g））。总酸度对乳酸菌计数影响最大。共分离鉴定出16 株乳酸菌：Lactobacillus fermentum（2 株）、Lactobacillus plantarum（3 株）、Lactobacillus brevis（2 株）、Lactobacillusacidophilus（1 株）、Lactobacillus casei（1 株）、Lactobacillus pentosus（1 株）、Lactobacillus delbrueckii（1 株）、Lactococcus lactis（3 株）、Pediococcus pentosaceus（2 株）。结论：眉山泡菜中乳酸菌存在多样性,实验结果为工业发酵生产泡菜积累了菌株。     

蒙古族奶嚼口与下层凝乳中乳酸菌的筛选和比较研究

该研究以蒙古族奶嚼口和下层凝乳为原料,对乳酸菌进行计数和分离,通过产酸凝乳、耐胆盐实验及基因分析筛选并鉴定优良发酵菌株。结果表明,奶嚼口和下层凝乳中乳酸菌活菌数为（12.60±0.78）lg（CFU/mL）和（12.31±0.35）lg（CFU/mL）;分离获得的200株乳酸菌中有19株产酸凝乳能力较好,其中12株来源于奶嚼口,7株来自下层凝乳;在0.3 g/L和0.6 g/L胆盐环境中,奶嚼口中筛选出的乳酸菌具有更强的胆盐耐受性;奶嚼口筛选出11株乳酸乳球菌（Lactococcus lactis）和1株屎肠球菌（Enterococcus faecium）;下层凝乳筛选出的7株均为乳酸乳球菌。奶嚼口和下层凝乳中乳酸菌活菌数及种属并无明显差异,奶嚼口中分离的具有产酸凝乳能力的乳酸菌多于下层凝乳中所筛选的菌株,并有较强的胆盐耐受性。     

D-lactic acid production by metabolically engineered Saccharomyces cerevisiae

Poly D-lactic acid is an important polymer because it improves the thermostability of poly L-lactic acid by the stereo complex formation. We constructed a metabolically engineered Saccharomyces cerevisiae that produces D-lactic acid efficiently. In this recombinant, the coding region of pyruvate decarboxylase 1 (PDC1) was completely deleted, and two copies of the D-lactate dehydrogenase (D-LDH) gene from Leuconostoc mesenteroides subsp. mesenteroides strain NBRC3426 were introduced into the genome. The D-lactate production reached 61.5 g/l, the amount of glucose being transformed into D-lactic acid being 61.2% under neutralizing conditions. Additionally, the yield of free D-lactic acid was also shown to be 53.0% under non-neutralizing conditions. It was confirmed that D-lactic acid of extremely high optical purity of 99.9% or higher. Our finding obtained the possibility of a new approach for pure d-lactic acid production without a neutralizing process compared with other techniques involving lactic acid bacteria and transgenic Escherichia coli.     

纯种乳酸菌接种发酵辣椒综合品质特性研究

为了获得高产γ-氨基丁酸（GABA）且综合性能优良的乳酸菌,以10株乳酸菌作为出发菌株对发酵辣椒进行纯种接种发酵,采用逼近理想解排序（TOPSIS）法分别从发酵产酸、抑菌性、亚硝酸盐积累以及生成γ-氨基丁酸能力等方面进行多因素综合评价。结果表明,发酵乳杆菌与食果糖乳杆菌的产酸速度较好,最终产酸量分别为0.020 8 g/100 g、0.020 3 g/100 g;乳链球菌和戊糖乳杆菌对大肠杆菌和金黄色葡萄球菌的抑菌作用最明显,其抑菌圈直径d=11 mm;乳酸片球菌产亚硝酸盐能力最弱,最终含量为1.7 mg/kg;食果糖乳杆菌与乳酸片球菌产GABA能力强,分别为1.97 mmol/L、1.86 mmol/L。采用TOPSIS法对10株菌种进行多因素综合评价,最终得到发酵乳杆菌与食果糖乳杆菌为综合品质特性较好的发酵优良菌株。     

Effect of added glucose and xylose on the fermentation of perennial ryegrass silage inoculated with Lactobacillus plantarum

Perennial ryegrass (Lolium perenne L) was ensiled in laboratory silos after addition of glucose or xylose at rates of 0, 25, 35 and 45 g kg^?1 fresh grass. In addition, an inoculum of Lactobacillus plantarum, supplying 10⁶ organisms g^?1 fresh grass, was applied to all treatments. Silos were opened after 7, 21 and 100 days and the silage was subjected to chemical and microbiological analysis. AH silages were well fermented with pHs between 3·60 and 3·70 and low NH₃-N concentrations (<95 g kg^?1 total nitrogen) and an absence of butyric acid. Glucose was virtually completely consumed within 21 days but 0·30–0·50 of the xylose doses remained after 100 days. Lactic acid concentrations were not increased by the addition of sugars, but the glucose treatments were associated with very high concentrations of ethanol, 60–100 g kg^?1 DM, and the xylose additions produced very high concentrations of acetic acid, 60–135 g kg^?1 DM. Most(>0·80) of the glucose that disappeared could be accounted for in ethanol formation but the xylose consumed could be accounted for only if the lactic acid produced in its fermentation was metabolised further to acetic acid; indeed, for the two higher doses of xylose, the concentrations of lactic acid were reduced from the control value of 177 g kg^?1 DM to 140 and 120 g kg^?1 DM, respectively. The results indicate that the provision of extra sugar, as hexose or pentose, allows yeasts to assume a more prominent role in the fermentation with consequent wasteful fermentation of sugars. Furthermore, the suggestion is that xylose may indirectly, via a stimulation of lactate-assimilating yeasts, encourage further metabolism of lactic acid to acetic acid.     

Efficient production of L‐lactic acid by Crabtree‐negative yeast Candida boidinii

Industrial production of L ‐lactic acid, which in polymerized form as poly‐lactic acid is widely used as a biodegradable plastic, has been attracting world‐wide attention. By genetic engineering we constructed a strain of the Crabtree‐negative yeast Candida boidinii that efficiently produced a large amount of L ‐lactic acid. The alcohol fermentation pathway of C. boidinii was altered by disruption of the PDC1 gene encoding pyruvate decarboxylase, resulting in an ethanol production that was reduced to 17% of the wild‐type strain. The alcohol fermentation pathway of the PDC1 deletion strain was then successfully utilized for the synthesis of L ‐lactic acid by placing the bovine L ‐lactate dehydrogenase‐encoding gene under the control of the PDC1 promoter by targeted integration. Optimizing the conditions for batch culture in a 5 l jar‐fermenter resulted in an L ‐lactic acid production reaching 85.9 g/l within 48 h. This productivity (1.79 g/l/h) is the highest thus far reported for L ‐lactic acid‐producing yeasts. DDBJ/EMBL/GenBank nucleotide database with Accession Nos. AB440630 and AB440631. Copyright © 2009 John Wiley & Sons, Ltd.     

Novel high butanol production from lactic acid and pentose by Clostridium saccharoperbutylacetonicum

We previously reported a butanol production process with pH-stat continuous feeding of dl-lactic acid and glucose as the co-substrate (Oshiro et al., Appl. Microbiol. Biotechnol., 87, 1177-1185, 2010). To accomplish butanol production from completely inedible substrates, in this study, we investigated acetone-butanol-ethanol (ABE) fermentation of Clostridium saccharoperbutylacetonicum N1-4 with lactic acid by using pentose as the co-substrate. Examination for optimum co-substrate indicated that arabinose was superior to glucose and xylose for ABE fermentation. Actually batch culture with lactic acid and arabinose without pH control exhibited higher butanol production (7.11?g/l) and lactic acid consumption (2.02?g) than those (6.62?g/l and 1.45?g, respectively) with glucose. Fed-batch culture without pH control increased these values to 12.08?g/l and 15.60?g/l butanol production, and to 3.83?g and 5.91?g lactic acid consumption by feeding the substrate once and twice, respectively. Finally, the result of gas chromatography-mass spectroscopy analysis using [1,2,3-(13)C(3)]-lactic acid indicated that lactic acid was converted to butanol with the efficiency of 51.9%. Thus, we established a novel high butanol production from lactic acid using arabinose as the co-substrate in simple fed-batch culture.     

风干武昌鱼中乳酸菌的分离鉴定及发酵性能研究

为获得具备优良性能的乳酸菌发酵剂,以我国极具特色的发酵肉制品—风干武昌鱼为基础原料,对传统风干武昌鱼在自然发酵过程中不同时期的乳酸菌进行分离鉴定与发酵性能测定。研究通过平板分离、菌落与菌体形态观察、糖醇发酵试验及生理生化试验鉴定所分离乳酸菌。结果表明所分离乳酸菌分属于7个种属:嗜盐四联球菌(Tetragenococcus halophilus)、弯曲乳杆菌(Lactobacillus curvatus)、乳酸乳球菌乳酸亚种(Lactococcus lactis subsp.lactis)、冷明串珠球菌(Leuconostoc gelidum)、格氏乳杆菌(Lactobacillus gasseri)、詹氏乳杆菌(Lactobacillus jensenii)和嗜淀粉乳杆菌(Lactobacillus amylohilus)。进一步采用产酸能力、食盐耐受性、亚硝酸盐耐受性、生长温度以及蛋白质和脂肪降解能力等试验测定菌株发酵性能,最终优化结果表明乳酸乳球菌乳酸亚种具备优良乳酸菌发酵剂的性能。