1株产耐热脂肪酶根霉突变株的选育

采用紫外线、硫酸二乙酯、紫外线和硫酸二乙酯复合对实验室保藏的1株高产耐热脂肪酶的根霉SFE-L01菌株进行诱变处理,筛选获得1株突变株UD-23,产耐热脂肪酶活力比出发菌株提高了112.39%,达24 000.00 U/mL。对突变株UD-23的产酶条件及部分酶学性质进行了研究,结果表明,突变株UD-23所产脂肪酶最适作用温度为60℃,且具有良好的热稳定性。条件优化后,该突变株产酶酶活达30 000.00 U/mL,比优化前提高了25%。     

米曲霉高产生淀粉酶菌株的诱变选育

以米曲霉y18为出发菌株,通过紫外诱变(UV)和硫酸二乙酯(DES)2次诱变,获得产生淀粉酶活力酶较高的菌株y18-U-36-D-40,酶活达到434.5U/mL,比出发菌株提高153.35%,菌株经过7次传代,酶活稳定。     

复合诱变皮状丝孢酵母选育高产油脂菌株

采用紫外线与硫酸二乙酯对皮状丝孢酵母进行复合诱变。研究结果表明:用15 W紫外灯在照射距离30 cm条件下照射60 s,可以筛选到油脂含量30.4%,生物量1.135 4 g/100 mL的突变菌株皮-60;利用硫酸二乙酯诱变皮-60,在硫酸二乙酯质量分数1.0%,处理时间为70 min条件下,可以筛选到油脂含量35.7%,生物量1.257 3 g/100 mL的突变菌株皮-60-70,较出发菌株,油脂含量提高了33.7%。     

豆豉纤溶酶产生菌的筛选及诱变

从豆豉中筛选出一株产纤溶酶活力为295.34 U/mL的菌株DXR25,经鉴定为芽孢杆菌属,DXR25经过紫外线、60Co、硫酸二乙酯(DES)、亚硝基胍(NTG)诱变,筛选得到突变株DXR7,其产酶活力为55.42.是出发菌株的2.2倍,最后对DXR7进行了10代的培养,结果表明,该突变株具有稳定的遗传性能.     

黑曲霉原生质体诱变及其产纤维素酶条件研究

采用紫外、硫酸二乙酯及复合诱变3种方法对黑曲霉原生质体进行诱变育种,以提高其酶活。结果表明,采用复合诱变效果最好,复合诱变所得菌株的酶活比出发菌株大幅度提高;应用单因素实验优化突变菌株的产酶条件,其最佳产酶条件培养基为:5%蔗糖、1.4%蛋白胨、0.2%磷酸二氢钾、0.05%硫酸镁,在250 mL摇瓶中装液60 mL,10%接种量、29℃下200 r/min培养96 h,酶活达到最大。     

硫酸二乙酯、紫外及~γCO_(60)辐射诱变筛选高产丁二酮菌株

以干酪乳杆菌亚种(Lactobacillus casei subsp.casei)ATCC393为出发菌株,其产丁二酮能力为57.31mg/L。分别采用硫酸二乙酯、紫外及γCO60辐射诱变育种,发酵,测丁二酮产量。γCO60辐射诱变后获得2株高产菌株,丁二酮产量分别为79.73mg/L和73.06mg/L,各为出发菌株的1.39倍和1.27倍。γCO60辐射诱变的最佳条件:50Gy/min剂量率,菌液浓度108个/mL,750Gy剂量照射处理。此时致死率为99.36%。     

普鲁兰酶产生菌的诱变育种

以从啤酒厂附近土壤中筛选得到的产偏碱性普鲁兰酶的细菌PUG12为出发菌株,对其进行紫外线和硫酸二乙酯复合诱变处理,从大量的突变株中筛选得到一株产普鲁兰酶活力较高的菌株YBC42,酶活力达8.32U/mL,比原出发菌株提高了3.4倍.     

高产溶纤酶菌株的选育

以枯草芽孢杆菌 YL-P2、YL-F2作为出发菌株,分别采用紫外线诱变、硫酸二乙酯诱变以及紫外线和硫酸二乙酯复合诱变,以脱脂牛奶平板法为初筛方法结合纤维蛋白平板复筛的方法选育出溶纤酶活力高的菌株, 获得了高产溶纤酶菌株 UV-8。UV-8溶纤酶活力达到2 314 IU/mL,与出发菌株相比,酶活力提高了2倍。结果表明,经多次传代后菌株 UV-8的溶纤酶酶活达到稳定,具有很好的潜在开发前景。     

紫外线和硫酸二乙酯诱变高产凝乳酶地衣芽孢杆菌的研究

以产凝乳酶地衣芽孢杆菌为出发菌株,通过紫外线诱变和硫酸二乙酯诱变,提高菌株的凝乳活力。经反复诱变筛选获得一株凝乳活力较高且水解活力较低的突变株DES-6,其凝乳活力为184.65SU/mL,比原菌株增加了15.41%,水解活力为23.35U/mL,比原菌株降低了64.80%。传代实验表明,突变株DES-6具有稳定的遗传性。     

产豆豉纤溶酶高产菌株的筛选及诱变育种

从豆豉中筛选出1株产纤溶酶的菌株DC33,鉴定为芽孢杆菌属,经过紫外线、^60Co、硫酸二乙酯（DES）、亚硝基呱（NTG）诱变筛选得到突变株DCDU7,其产酶活力为644．77U／mL,是出发菌株的2．4倍。对DCDU7菌株进行了10代的培养,结果表明该突变株具有稳定的遗传性能。     

Nisin release from films is affected by both protein type and film-forming method

Effects of protein type (wheat or corn) and film-forming method (casting or heat-pressing) on films were evaluated for the retention of biologically active nisin (Nisaplin) and release of activity into water at four different temperatures (5, 25, 35 and 45 °C). Nisin activity was measured using the agar diffusion method against Lactobacillus plantarum 1752. Cast corn zein (CCZ) and cast wheat gluten (CWG) films retained 12.1% (8.1×10⁴ IU/g film) and 15.8% (1.1×10⁵ IU/g film) of the original activity after film formation, respectively. Heat-pressed corn zein (HPCZ) and heat-pressed wheat gluten (HPWG) films retained 6.5% (4.3×10⁴ IU/g film) and 7.4% (4. 9×10⁴ IU/g film) of the original activity after film formation, respectively. The maximum nisin activity found migrating into water at any sampling time was 561 IU/ml (CCZ), 1058 IU/ml (CWG), 309 IU/ml (HPCZ), and 478 IU/ml (HPWG).     

密码子优化及透明颤菌血红蛋白共表达提高耐热脂肪酶在毕赤酵母的表达

根据毕赤酵母密码子的偏好性,通过在线软件对Thermomyces lanuginosus脂肪酶基因（tll）和透明颤菌血红蛋白基因（vhb）进行密码子优化。优化后的脂肪酶基因（tll-opt）和透明颤菌血红蛋白基因（vhb-opt）中碱基GC含量分别由原来的46%提高到49%和42%提高到51%,碱基A、T、C、G均匀分布,减少了AT和GC富集区,利于tll-opt和vhb-opt基因在毕赤酵母X33中的表达。将tll和tll-opt连接到表达载体pPICZαA并转入毕赤酵母X33中。摇瓶培养条件下,含有tll和tll-opt重组工程菌的最大酶活力分别为7 U/mL和16 U/mL。50 L发酵罐培养条件下,含有tll和tll-opt重组工程菌的最大酶活力分别为201 U/mL和430 U/mL。为了进一步提高含tll-opt重组工程菌的表达酶活性,将vhb-opt转入该重组工程菌得到工程菌VHb＋（含有tll-opt和vhb-opt）。重组工程菌VHb＋在摇瓶培养条件下最大酶活力为23 U/mL,分别是优化后基因和原始基因最大表达酶活力的1.39 倍和3.28 倍。重组工程菌VHb＋在50 L发酵罐瓶培养条件下最大酶活力为610 U/mL,分别是优化后基因和原始基因最大表达酶活力的1.41 倍和3.03 倍。     

Inhibition of Listeria monocytogenes on the surface of individually packaged hot dogs with a packaging film coating containing nisin

The objective of this study was to determine the effectiveness of packaging films coated with a methylcellulose/hydroxypropyl methylcellulose-based solution containing 10,000, 7,500, 2,500, or 156.3 IU/ml nisin for controlling Listeria monocytogenes on the surfaces of vacuum-packaged hot dogs. Barrier film coated with a methylcellulose/hydroxypropyl methylcellulose-based solution containing nisin or no nisin (control) was heat sealed to form individual pouches. Hot dogs were placed in control and nisin-containing pouches and inoculated with a five-strain L. monocytogenes cocktail (approximately 5 log CFU per package), vacuum sealed, and stored for intervals of 2 h and 7, 15, 21, 28, and 60 d at 4 degrees C. After storage, hot dogs and packages were rinsed with 0.1% peptone water. Diluent was spiral plated on modified oxford agar and tryptic soy agar and incubated to obtain counts (CFU per package). L. monocytogenes counts on hot dogs packaged in films coated with 156.3 IU/ml nisin decreased slightly (approximately 0.5-log reduction) through day 15 of refrigerated storage but was statistically the same (P > 0.05) as hot dogs packaged in films without nisin after 60 d of storage. Packaging films coated with a cellulose-based solution containing 10,000 and 7,500 IU/ml nisin significantly decreased (P < 0.05) L. monocytogenes populations on the surface of hot dogs by greater than 2 log CFU per package throughout the 60-d study. Similar results were observed for hot dogs packaged in films coated with 2,500 IU/ml nisin; however, L. monocytogenes populations were observed to be approximately 4 log CFU per package after 60 d of refrigerated storage from plate counts on tryptic soy and modified oxford agars.     

产ε-聚赖氨酸白色链霉菌的化学诱变育种

该研究以白色链霉菌（Streptomyces albus）BNCC 186223为原始菌株,使用硫酸二乙酯对其进行化学诱变,考察最佳诱变条 件,并筛选高产ε-聚赖氨酸的菌株。结果表明,最佳诱变条件为硫酸二乙酯10 μL/mL,诱变时间45 min。在此最佳诱变条件下,获得一株 高产ε-聚赖氨酸突变菌株DES-27,其ε-聚赖氨酸产量为2.90 g/L,较未诱变前提高38.10%。 因此,硫酸二乙酯的诱变作用能够显著提 高ε-聚赖氨酸的产量。     

海洋纤溶酶高产菌株的选育及产酶条件优化

以海洋枯草芽孢杆菌FA-7为出发菌株,经紫外诱变获得一株遗传稳定性良好的高产纤溶酶菌株Y-22,并对该菌株的发酵培养基和发酵条件进行优化,确定菌株Y-22的最佳产酶条件为可溶性淀粉4％,黄豆粕粉2．5％,CaCl20．025％,MgSO4·7H：00．35％,吐温-80O．15％;接种量4％,装液量50mt：250mL,初始pH值为5．5,发酵温度30℃,180r／min振荡培养96h。在此条件下,菌株Y-22的发酵液粗酶酶活为（910±11．3）1U／mL,是出发菌株的3．05倍。     

Synergistic effect of nisin and heat treatment on the growth of Escherichia coli O157:H7

A combination of nisin and heat treatment was found to inhibit Escherichia coli O157:H7 effectively. After organisms were heated at 50, 52.5, and 55 degrees C for 5, 10, and 15 min, respectively, nisin was incorporated into the plates of E. coli O157:H7 at 0, 25, 50, and 100 IU/ml. The concentration of 100 IU/ml nisin significantly inhibited the growth of E. coli O157:H7 heated at 50 and 52.5 degrees C for 15 min. Nisin treatment at 100 IU/ml for 6 h resulted in the elimination of E. coli O157:H7 heated at 55 degrees C for 10 and 15 min.     

培养基与分割发酵优化促进Nisin生物合成

为了提高乳酸乳球菌（Lactococcus lactis）细胞生长和乳酸链球菌素（Nisin）的合成效率,以正交优化法研究培养基分批发酵和分割发酵方式对Nisin生物合成效率的影响。结果表明,优化发酵培养基配方为：5%蛋白胨,3%蔗糖,2%玉米浆,1%酵母浸粉。在此优化条件下,摇瓶培养（11 h）峰值生物量为4.9×109 CFU/mL,较对照提高43%;10 L发酵罐分批发酵峰值生物量、Nisin效价及Nisin合成速率■q分别为7.75×109 CFU/mL、2 573 IU/mL、151.4 IU/（mL·h）,较优化前分别提升38.0%、56.6%、38.2%。分批发酵培养18 h Nisin达到峰值后[■q为147 IU/（mL·h）],以不同比例分割发酵并继续培养7 h,第二次分割（25%）为Nisin合成最适发酵方式,其Nisin平均合成速率达到294 IU/（mL·h）,较分批发酵提高了100%。     

Effect of thaw rates on survival of buffalo spermatozoa frozen straws

Eighteen ejaculates from three buffalo bulls of Nili-Ravi breed were tested in a 3 X 6 X 3 factorial experiment. Semen was extended in lactose-fructose-egg yolk-glycerol extender containing penicillin (1000 IU/ml) and streptomycin (1000 micrograms/ml). Semen was frozen in .5-ml polyvinyl chloride straws in liquid nitrogen vapor and stored in liquid nitrogen for 24 h. Straws were thawed at water bath temperatures of 0, 37, or 75 degrees C for 2 min, 15 s, and 9 s, respectively. At thawing bath temperature of 0, 37, or 75 degrees C, percentage of motile spermatozoa averaged 30, 40, and 50%. Differences were significant between thaw rates for initial postthaw motility, postthaw sperm survival at 37 degrees C, and absolute index of survival of spermatozoa. Bulls were also different for initial postthaw motility, postthaw sperm survival at 37 degrees C, and absolute index of survival of spermatozoa. Thaw rate of 75 degrees C for 9 s was superior to other rates.     

酶法提取麒麟菜膳食纤维工艺的研究

分别采用正交和均匀设计法,优选出麒麟菜(Eucheuma)膳食纤维的最佳漂白和提取工艺条件,并对提取的膳食纤维进行了分析。结果表明:最佳漂白条件--漂白液浓度2g/L、pH7.0、漂白时间40min;最佳提取条件--加水量4%(L/g)、煮沸时间60min、在60～65℃下分别加入0.15%的蛋白酶和0.1%的α-淀粉酶、分别酶解60和30min。在此条件下,提取率为39.06%,膨胀力为38.6ml/g、持水力为2045.8%,膳食纤维含有较低的蛋白质、脂肪且无淀粉检出,具有较高含量的Ca、P、K等矿物元素,其功能指标优于化学法提取的麒麟菜膳食纤维和小麦麸皮标准膳食纤维,有望在人体中发挥重要的生理功效。