枯草芽胞杆菌与酵母菌混合发酵奶牛精饲料的研究

对混菌发酵奶牛精饲料的发酵条件进行了研究。采用枯草芽孢杆菌与酵母菌混合发酵的方法,研究菌种比例、麸皮含量、水分、腐植酸钠添加量对发酵产物中淀粉酶和中性蛋白酶含量的影响。结果表明:奶牛精饲料的最佳发酵条件为:枯草芽孢杆菌与酵母菌的接种比例为3∶7,麸皮添加量为5%,水分为65%,腐植酸钠在种子液中的添加量为2%,在发酵底物中的添加量为2%。在此条件下发酵奶牛精饲料,测得产品中中性蛋白酶含量为1 476.00U/g,淀粉酶含量为6 339.24U/g,与未发酵精饲料相比,中性蛋白酶含量是未发酵精料的3倍,淀粉酶含量是未发酵精料的1.52倍。     

枯草芽孢杆菌ZC1发酵产中性蛋白酶的条件优化

在摇瓶发酵条件下,对枯草芽孢杆菌产中性蛋白酶条件进行优化。在单因素的基础上,通过Plackett-Burman实验和最徒爬坡实验确定出响应面实验设计的中心点:豆粕粉66g/L,麦芽糖16g/L及磷酸二氢钾8.5g/L。通过响应面实验设计,得到最佳发酵条件为:豆粕粉67.13g/L,麦芽糖16.17g/L,磷酸二氢钾8.52g/L,吐温-80 3mL/L,海藻糖3g/L,pH 7.5,接种量4%,温度37℃。在该发酵条件下发酵72h,中性蛋白酶酶活由原来的454.2U/mL提高到993.1U/mL。     

产蛋白酶菌株的鉴定及其对虾壳的脱蛋白研究

从虾塘沉积物中分离到一株产蛋白酶的菌株C1,采用菌落形态、生理生化特征和16S rDNA基因序列分析相结合的方法进行鉴定,进一步探究其在提取虾壳甲壳素工艺中脱蛋白的应用,并与枯草芽孢杆菌（Bacillus subtilis）对虾壳蛋白脱除效果进行比较分析。结果表明,菌株C1被鉴定为一株蜡样芽孢杆菌（Bacillus cereus）,其对虾壳的脱蛋白能力高于枯草芽孢杆菌。当发酵培养基中葡萄糖添加量为50 g/L,虾壳粉添加量为20 g/L,酵母膏添加量为1 g/L时,枯草芽孢杆菌和蜡样芽孢杆菌发酵5 d的蛋白酶活力分别为145.7 U/mL、220.8 U/mL,脱蛋白率分别达到80.4%、90.8%。     

芽孢杆菌的产酶特性及其对抗生素的耐受性

以4株枯草芽孢杆菌（Bacillus subtilis）和2株地衣芽孢杆菌（Bacillus licheniformis）为研究对象,通过测定其成胞率、中性蛋白酶活性及对24种抗生素的耐受性,从中筛选优良芽孢杆菌。结果表明,从6株芽孢杆菌中筛选得到3株成胞率和产中性蛋白酶均较好的芽孢杆菌,分别为枯草芽孢杆菌BLCC1-0552、BLCC1-0615和地衣芽孢杆菌BLCC1-0441,液体发酵24 h后,成胞率较高,均达到98%;中性蛋白酶活性分别为90.58 U/mL、100.32 U/mL和24.72 U/mL。其中,枯草芽孢杆菌BLCC1-0615固体发酵玉米-豆粕型常规饲料产中性蛋白酶活力最高,发酵48 h时达到2 154.49 U/g。3株芽孢杆菌对抗生素的耐受性不一致,其中枯草芽孢杆菌BLCC1-0615对24种抗生素中的17种表现敏感,敏感率最高为70.83%。因此,确定优良菌株为枯草芽孢杆菌BLCC1-0615,其成胞率、产中性蛋白酶能力好及对抗生素耐受性较低,具有作为饲料添加剂应用于畜禽饲料中的潜力。     

凝结芽孢杆菌高密度发酵及产孢关键因素研究

以凝结芽孢杆菌TZB-5菌株为研究对象,在高密度发酵及提高产孢率两方面展开探究,旨在为其微生态制剂的制备提供参考。结果表明:麸皮浸出液能大幅度提高菌密度,硫酸铁是促进细胞形成芽孢的关键因素。菌株高密度发酵最佳培养基配方:葡萄糖15 g/L、酵母粉5 g/L、氯化钠5 g/L、麸皮浸出液80%、pH5.5;最佳培养条件:培养温度47 ℃、转速200 r/min、培养时间20 h,在此条件下发酵的菌密度高达5.5×10~9 cfu/mL;菌株产孢最佳培养基配方:可溶性淀粉10 g/L、豆粕粉10 g/L、酵母粉5 g/L、氯化钠5 g/L、硫酸铁100 mg/L、麸皮浸出液80%、pH5.5,芽孢产量最高达到5.1×10~9 cfu/mL,芽孢率为98.15%。     

一株饲用益生枯草芽孢杆菌液体发酵的研究

为解决工业化生产中益生菌枯草芽孢杆菌(Bacillus subtilis)活菌数量低、成本高的问题,根据摇瓶正交试验结果对枯草芽孢杆菌的液体发酵条件进行了深入研究,结果表明:最佳培养基配方为玉米粉100g/L,豆粕粉25g/L,硫酸锰30mg/L,硫酸镁1g/L,磷酸二氢钠1.5g/L,磷酸氢二钠3g/L;最优发酵工艺条件为培养温度30℃,初始pH 7.0,通气量2.0m3/h,罐压0.03MPa,发酵周期36h。细菌浓度达到了8.0×1010cfu/mL,芽孢形成率达到94%。     

山西老陈醋源优良芽孢杆菌菌株的鉴定及筛选

以分离自山西老陈醋醋醅中的9株芽孢杆菌为研究对象,对菌株进行分子生物学鉴定,并对其产酶、产酸、产酯、产多酚和产乙偶姻等发酵特性综合研究。结果表明,9株芽孢菌分别被鉴定为地衣芽孢杆菌（Bacillus licheniformis）CP-18、CP-1853、CP-1671,枯草芽孢杆菌（Bacillus subtilis）CP-803,解淀粉芽孢杆菌（Bacillus amyloliquefaciens）CP-2430,甲基营养型芽孢杆菌（Bacillus methylotrophicus）CP-6、CP-1576,漠海威芽孢杆菌（Bacillus mojavensis）CP-15和球形赖氨酸芽孢杆菌（Lysinibacillus sphaericus）CP-2436。9株菌的淀粉酶、纤维素酶和蛋白酶酶活分别为9.77～61.40 U/mL、7.98～54.99 U/mL和4.16～22.80 U/mL,其中菌株CP-1853的蛋白酶活最高（22.80 U/mL）,且具有较高淀粉酶活（43.22 U/mL）和纤维素酶活（38.97 U/mL）;菌株CP-18 和CP-1853产酸和产酯能力较强,分别为0.25 g/L和10.67 g/100 mL;菌株CP-803和CP-1576产多酚和乙偶姻能力较强,分别为596.38 μg/mL和1.15 mg/mL。这些优良菌株可以为山西老陈醋的提质增效起到一定强化作用。     

富含低聚木糖的小麦麸皮饮料的研制

以小麦麸皮为原料,经焙烤、酶解、澄清和调配等工艺制成富含低聚木糖的保健功能性饮料。先将小麦麸皮在180℃下焙烤20 min,之后采用木聚糖酶将其中的木聚糖酶解为低聚木糖,采用风味蛋白酶将蛋白质酶解为短肽,中温α-淀粉酶将淀粉酶解为葡萄糖和麦芽糖。优化后的酶解条件为:木聚糖酶153 U/g麸皮、风味蛋白酶138 U/g麸皮、中温α-淀粉酶60 U/g麸皮,料水比1∶8(g∶mL),pH值6.0,反应温度50℃,反应时间4 h。酶解液经稀释后加入0.2 g/L皂土和0.1 g/L壳聚糖澄清,调配时加入10 g/L蜂蜜、60 g/L白砂糖和0.75 g/L柠檬酸,经超高温瞬时杀菌、无菌灌装得到成品。低聚木糖(2.06 mg/mL)和短肽为饮料中主要功能性成分。     

脉冲磁场诱变结合发酵条件优化提高 中性蛋白酶产量的研究

利用脉冲磁场诱变枯草芽孢杆菌结合发酵条件优化以实现中性蛋白酶产量的最大化。结果表明在磁场强度为7 T,脉冲数为40次时诱变得到了中性蛋白酶产量提高17.7%的枯草芽孢杆菌菌株。对其发酵条件进行优化,得到最佳发酵培养基配方为豆粕粉90 g/L,麦芽糖10 g/L,KH_2PO_4 8 g/L,最佳发酵条件为接种量5%、发酵温度37℃、摇床转速180 r/min。在此条件下,发酵48 h,中性蛋白酶酶活力为384.57 U/m L,比野生株提高了26.48%。     

枯草芽孢杆菌L1发酵产中性蛋白酶活性的研究

中性蛋白酶在工业上具有广泛的应用,文中以豆粕为主要氮源,探讨了枯草芽孢杆菌L1菌株发酵豆粕产中性蛋白酶的酶活性。在单因素试验的基础上,通过正交试验优化了枯草芽孢杆菌L1产中性蛋白酶的发酵条件,即豆粕粉浓度为2.0%,葡萄糖为1.0%,接种量为7%,发酵温度为36℃,其发酵液的中性蛋白酶活力可达68.7U/m L。     

枯草芽孢杆菌生产抗菌物质的食品级发酵培养基优化

为得到安全可靠的新型生物防腐剂,用枯草芽孢杆菌发酵食品同时提高其抗菌活性。采用单因素试验和Plackett-Burman试验设计筛选出4 个关键因子为脱脂奶粉、番茄汁、发酵时间和发酵温度;使用Box-Behnken原理设计进行响应面试验确定出最佳培养基配方和发酵条件：小麦粉16.0 g/L、脱脂奶粉20.0 g/L、黄豆粉酶解液200.0 mL/L、番茄汁40.0 mL/L、NaCl 10.0 g/L,装液量50.0 mL、发酵温度32.14 ℃、发酵时间60.0 h。拟合实验模型结果显示：优化后发酵液对荧光假单孢的抑菌圈直径较优化前提高42%,对短小芽孢杆菌的抑菌圈直径较优化前提高47%。     

Antimicrobial activity of epsilon-polylysine against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in various food extracts

ABSTRACT:  This study compared the antimicrobial effects of ɛ-polylysine (ɛ-PL) against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in 6 food extracts and in broth. The food extracts (10% (w/w) in distilled water) evaluated were fat-free and whole fat milk, beef, bologna, rice, and vegetables (50:50 ratio of broccoli and cauliflower). ɛ-PL was tested at 0.005% and 0.02% (w/v) against E. coli O157:H7 and L. monocytogenes , and 0.02% and 0.04% (w/v) against S. Typhimurium . The substrates were inoculated (5 log CFU/mL) and periodically analyzed for surviving populations during storage at 12 °C for 6 d. In general, all 3 pathogens reached 7 to 9 log CFU/mL within 2 d in control substrates (no ɛ-PL). Immediate bactericidal effects ( P < 0.05) following exposure to ɛ-PL were obtained in the rice (all pathogens) and vegetable ( E. coli O157:H7 and S. Typhimurium ) extracts. During storage, antimicrobial effects of ɛ-PL were more pronounced in the food extracts than in the broth medium. The greatest antimicrobial activity for all 3 pathogens was obtained in the rice and vegetable extracts, where counts were reduced ( P < 0.05) to below the detection limit (0.0 log CFU/mL) by one or both ɛ-PL concentrations tested. In the other food extracts (fat-free milk, whole fat milk, beef, and bologna), both ɛ-PL concentrations tested generally resulted in lower ( P < 0.05) pathogen levels at the end of storage compared to initial counts, with better bactericidal effects exerted by the higher of the 2 ɛ-PL concentrations. Additional research is needed to explore the potential antimicrobial effects of ɛ-PL in real food systems.     

制备鲐鱼鱼肉发酵液中抗氧化因子的条件优化

以食品级枯草芽孢杆菌为实验用菌,通过发酵法制备具有抗氧化作用的鲐鱼鱼肉发酵液。在液体培养基的基础上研究装液量、葡萄糖添加量、鱼肉培养基料液比、培养转速对发酵产物的影响,结果表明：装液量50mL/250mL或100mL/250mL、葡萄糖添加量2%、鱼肉:水料液比(m/V)1:(1～2)、摇床转速150r/min条件下,发酵液的抗氧化性较高。在加糖量、装液量及料液比对试验结果影响较大的单因素试验基础上,采用响应面分析法(Box-Behnken)对发酵鱼肉培养基制备抗氧化型发酵液的工艺参数进行优化后,得出最佳条件为：葡萄糖添加量3.98%、装液量96.02mL/250mL、鱼肉:水料液比1:1.60,其发酵液的DPPH自由基清除率可达 94.52%。     

好氧堆肥用枯草芽孢杆菌GX2产芽孢工艺优化

芽孢杆菌是好氧堆肥微生物菌剂的重要组成部分,为降低菌剂生产成本、改善其品质及应用效果,对芽孢杆菌产芽孢工艺进行研究.首先,在摇瓶上进行枯草芽孢杆菌GX2的培养基及培养条件优化,在此基础上,在3L发酵罐中进行分批补料优化研究,进一步提高细胞浓度,最后,通过单因素及正交试验研究,在3 L发酵罐中对产芽孢工艺进行了优化.结果...     

Bacillus stearothermophilus麦芽糖淀粉酶在Bacillus subtilis中的重组表达及发酵优化

为了实现Bacillus stearothermophilus嗜热脂肪芽孢杆菌来源的麦芽糖淀粉酶基因amyM(EC 3.2.1.133)在枯草芽孢杆菌中的重组表达,以opt-amyM/T质粒为模板进行PCR扩增得到目的基因,与表达载体pHY300PLK进行重组连接后转入宿主菌Bacillus subtilis CCTCC M 2016536中进行表达。在TB培养基中发酵培养48 h,麦芽糖淀粉酶酶活达到250.7 U/mL。继续对重组菌氮源种类及复配、氮源质量浓度、碳源质量浓度等摇瓶发酵条件进行优化,确定其最佳产酶条件为:复合氮源为酵母浸膏25 g/L和大豆蛋白胨5 g/L、葡萄糖5 g/L、培养基初始pH 6.5、最适培养温度41℃;在此条件下麦芽糖淀粉酶酶活可达396.7 U/mL,是优化前的1.6倍。在此基础上进一步对麦芽糖淀粉酶进行酶学性质进行测定:麦芽糖淀粉酶最适温度为60℃,最适pH为5.5,半衰期为325 h,Km为0.95 g/L,比活为2646 U/mg。     

Production of polysaccharide and surfactin by Bacillus subtilis ATCC 6633 using rehydrated whey powder as the fermentation medium

The aim of this research was to assess the amounts of polysaccharide and surfactin produced by Bacillus subtilis ATCC 6633 in rehydrated whey powder (RWP) as the growth medium. One-day-old cultures of B. subtilis (～4.6 log cfu/mL) were inoculated into 100mL of 10, 15, or 20% (wt/vol) RWP and incubated at 30°C for 72 h. To analyze the effects of lactose and protein on polysaccharide and surfactin production, 6 RWP solutions containing different levels of lactose and protein were also used as media. The number of vegetative cells and spores, pH, viscosity, and the concentration of lactose were determined at 0, 24, 48, or 72 h of fermentation. The levels of polysaccharide and surfactin produced after 72 h of fermentation were measured using HPLC and the phenol-sulfuric acid method, respectively. During 72 h of fermentation, B. subtilis populations increased from 4.6 to 10.54, 9.82, and 9.67 log(10) cfu/mL in 10, 15, and 20% RWP, respectively. The number of B. subtilis spores in 10% RWP increased from 3.91 to 4.72 log(10) cfu/mL after 48 and 72 h of fermentation, respectively. The increased level of lactose or protein in RWP did not significantly change the vegetative growth. After 72h of fermentation, the pH of RWP decreased from 5.70 to 4.99 with a slight increase in viscosity. Polysaccharide levels in 10, 15, and 20% RWP after fermentation were 513.6, 613.5, and 768.3mg/L, respectively, with B. subtilis producing 0.18 to 0.29 g/L of surfactin after 72 h of fermentation. The polysaccharide or surfactin production was not changed significantly by addition of protein or lactose to RWP. These results indicate that RWP is a good fermentation substrate for surfactin and polysaccharide production.     

产谷氨酸脱羧酶重组枯草芽孢杆菌的构建与食品级表达

谷氨酸脱羧酶(GAD)是生物合成γ-氨基丁酸(GABA)的关键酶。本研究构建了4株无抗标记的重组枯草芽孢杆菌,首次实现了乳酸乳球菌(Lactococcus lactis ssp.lactis IL 1403)来源的谷氨酸脱羧酶基因在枯草芽孢杆菌中的食品级表达。通过比较4株重组枯草芽孢杆菌的生长曲线和发酵酶活曲线,筛选出产酶效率最高的重组菌株B.subtilis WB600/pUB-P43-gadB(opt)-dal,该重组菌在初始发酵培养基中发酵42 h后发酵酶活可达4.1 U/mL。通过调整培养基成分,重组菌B.subtilis WB600/pUB-P43-gadB(opt)-dal的发酵酶活最高达到7.4 U/mL,与初始相比酶活提高了79%,是目前已报道重组枯草芽孢杆菌产谷氨酸脱羧酶酶活的最高水平。     

Antilisterial Activity of Hops Beta Acids in Broth with or Without Other Antimicrobials

ABSTRACT: Hops beta acids (HBA) are parts of hops flowers used to preserve wort and provide flavor in beer, and are reported as having antimicrobial properties. This study evaluated the antilisterial activity of HBA alone or in combination with other known antimicrobials in a culture broth medium. Listeria monocytogenes (10‐strain mixture) was inoculated (2.6 to 2.8 log CFU/mL) into tryptic soy broth supplemented with 0.6% yeast extract (TSBYE) without (control) or with HBA (0.5 to 5.0 μg/mL), potassium lactate (1.0%), sodium diacetate (0.25%), or acetic acid (0.1%), alone or in combination with HBA (0.5 to 3.0 μg/mL). Survival/growth of the pathogen during storage at 4 °C (35 d), 10 °C (20 d), or 25 °C (2 d) was periodically monitored by spiral plating onto tryptic soy agar plus 0.6% yeast extract. As expected, TSBYE without antimicrobials (control) supported rapid pathogen growth with growth rates of 0.40, 2.88, and 9.58 log CFU/mL/d at 4, 10, and 25 °C, respectively; corresponding Y_end values exceeded 9.0 log CFU/mL at 35, 20, and 2 d storage. HBA used alone (1.0 to 5.0 μg/mL) inhibited growth of L. monocytogenes at all 3 temperatures, with inhibition being more pronounced at higher concentrations and at the lower storage temperature (4 °C). The antilisterial activity of HBA (0.5 to 3.0 μg/mL) was enhanced when combined with sodium diacetate, acetic acid, or potassium lactate, achieving complete inhibition at 4 °C when 3.0 μg/mL HBA were used in combination with each of the above antimicrobials. Overall, HBA exhibited promising antilisterial activity in a broth medium and further studies are needed to investigate its potential antilisterial effects in food products.     

纳豆激酶液态发酵工艺优化

在单因素实验的基础上,用响应面法和正交实验对Bacillus subtilis液态发酵生产纳豆激酶的培养基和发酵条件进行了优化。实验结果表明,最佳产酶培养基组成为麦芽糖2%,酵母膏4%,CaCl20.03%,pH7.0;最佳发酵条件为接种量1%,培养温度34℃,摇床转速200r/min,装液量100mL/500mL(挡板瓶),培养48h达到产酶高峰,产酶活力可达4300U/mL。     

里氏木霉(Trichoderma reesei)306产组织型纤溶酶原激活剂(t-PA)液体发酵条件的优化

采用正交设计和响应面分析等实验方法对基因工程菌株里氏木霉 (Trichodermareesei)30 6产组织型纤溶酶原激活剂 (t PA)的液体发酵条件进行了优化。确定了适宜的培养基配方和最佳发酵工艺条件。在优化发酵条件下 ,摇瓶液体发酵液中的t PA酶活力达 3386 91IU/mL ,比初始发酵条件下酶活力提高上千倍。