臭氧对耐酸耐热菌作用的研究

以耐酸耐热菌为研究对象,研究了臭氧对它的致死作用,并观察了臭氧作用时间、温度、pH等对臭氧杀菌效果的影响,为果汁行业实际应用臭氧杀菌提供实验依据。      

臭氧对耐酸耐热菌作用的研究

以耐酸耐热菌为研究对象,研究了臭氧对它的致死作用,并观察了臭氧作用时间、温度、pH等对臭氧杀菌效果的影响,为果汁行业实际应用臭氧杀菌提供实验依据。     

PCR技术检测沙门氏菌反应条件的优化

沙门氏菌是引起细菌性食物中毒的重要病源之一,因此建立一种快速、简便、灵敏的检测方法十分重要.该研究以沙门氏菌的侵袭蛋白基因inVA为靶细胞设计引物,进行PCR扩增并对PCR反应体系中引物量、模板量、dNTP、退火温度和PCR循环数等进行优化,以确定适宜的PCR反应体系和PCR扩增反应程序.     

PCR法快速检测耐热霉菌雪白丝衣霉

以耐热性霉菌——雪白丝衣霉为研究对象,根据Gen-bank公布的beta-tubulin基因序列,设计两对引物,经过PCR筛选,确定引物Bn-1用于建立雪白丝衣霉的PCR检测方法。对PCR法的扩增条件退火温度和引物浓度进行优化,59℃的退火温度和0.2μM/PCR反应体系的引物浓度为最佳条件;考察该PCR方法的灵敏度和特异性,基因组DNA的灵敏度为1ng/PCR反应体系,检测特异性好。该方法为雪白丝衣霉的快速检测提供了一种重要的技术手段。     

耐热过氧化氢酶基因工程菌的构建及其发酵条件

利用PCR扩增技术以嗜热脂肪芽孢杆菌IAM11001染色体DNA为模板,扩增得到嗜热脂肪芽孢杆菌过氧化氢酶编码基因perA。再与经EcoRⅠ酶切的温控表达载体pBV220连接,构建重组质粒,并转化宿主大肠杆菌JM109,得到耐热过氧化氢酶基因工程菌,然后对该工程菌在LB培养基和半合成培养基中的发酵条件进行了优化。结果表明：在LB培养基中,诱导时期和酶合成最佳诱导时间均为5h,最大产酶量达到72．9U／mL;在半合成培养基中,于30℃培养4h,再于42℃诱导培养6h,产酶量最高达到131U／mL。     

苹果汁中耐热菌培养基的优化及生长曲线

通过对苹果汁中分离耐热菌和德国标准耐热菌菌株基础培养基的正交优化,确定最佳的培养基。得出最佳培养基配比是(质量分数):德国标准菌:蛋白胨0.5%,果糖0.2%,酵母粉0.2%,MgSO4·2H2O0.1%,CaCl20.5%,KH2PO40.12%,MnSO4·4H2O0.05%;分离菌:蛋白胨0.5%,葡萄糖0.2%,酵母粉0.2%,MgSO4·2H2O0.1%,CaCl20.5%,KH2PO40.12%,MnSO4·4H2O0.05%。并计算得出吸光度与细菌浓度之间的关系公式及用优化培养基培养绘制耐热菌生长曲线。     

苹果汁中耐热菌培养基的优化及生长曲线

通过对苹果汁中分离耐热菌和德国标准耐热菌菌株基础培养基的正交优化,确定最佳的培养基。得出最佳培养基配比是(质量分数):德国标准菌:蛋白胨0.5%,果糖0.2%,酵母粉0.2%,MgSO4·2H2O0.1%,CaCl20.5%,KH2PO40.12%,MnSO4·4H2O0.05%;分离菌:蛋白胨0.5%,葡萄糖0.2%,酵母粉0.2%,MgSO4·2H2O0.1%,CaCl20.5%,KH2PO40.12%,MnSO4·4H2O0.05%。并计算得出吸光度与细菌浓度之间的关系公式及用优化培养基培养绘制耐热菌生长曲线。      

耐热纤维素酶产酶条件的优化

在原始发酵培养条件下,枯草芽孢杆菌的酶活力达到1.0U/ml。对影响其产酶水平的培养基中的碳源、氮源、无机盐等发酵培养条件进行了单因子试验研究,并对碳、氮、磷三因素进行正交试验,对原始培养基进行了优化,结果在CMC-Na1.5%,酵母粉1.0%,K2HPO 40.15%,NaCl0.5%,MgSO 40.05%,蛋白胨1.5%,初始pH为7.0的优化培养基中,于37℃,200转/分的转速下,培养48小时,产生的纤维素酶活力达到1.5U/ml。     

双重PCR检测黄秋葵储藏期间青霉和链格孢菌条件的优化

以黄秋葵为材料,分离纯化在贮藏过程中引起腐烂的主要病原真菌青霉和链格孢菌。根据基因库上的序列,针对青霉和链格孢菌在18SRNA的序列设计2对引物,建立了用于检测黄秋葵青霉和链格孢菌双重PCR检测方法,并对PCR扩增条件进行了优化,优化条件如下:引物Alt4、Alt5和Pen F'、Pen R的比例为2∶1、退火温度为52℃、Mg~(2+)浓度为2.5 mmol/L、d NTP浓度为0.08 mmol/L、循环数为35、延伸时间为50 s、退火时间为30 s。结果表明,该方法灵敏度较高、特异性强。该试验可以为黄秋葵在储藏期间的病害防治提供重要指导依据。     

Inhibition of Alicyclobacillus acidoterrestris spores by natural compounds

The aim of this study was to evaluate the antimicrobial effectiveness of some natural compounds (cinnamaldehyde, eugenol, limonene) and sodium benzoate against two strains of Alicyclobacillus acidoterrestris (c8 and γ4). The antimicrobial compounds (10–500 ppm) were solved in malt extract broth, inoculated separately with 10³ spores mL⁻¹ of each strain; the samples were incubated at 44 °C and the outgrowth of spores was evaluated every day by measuring the absorbance of the medium at 420 nm; inoculated samples without active compounds were used as controls. The results pointed out that limonene was not effective in inhibiting the outgrowth of A. acidoterrestris spores; 100 ppm of cinnamaldehyde or sodium benzoate slowed the spore germination, whereas 500 ppm of eugenol inhibited the growth of microbial targets for 13 days. Strain c8 was more resistant than isolate γ4 and cinnamaldehyde was the most effective compound in inhibiting the germination of A. acidoterrestris spores.     

Thermal resistance of Alicyclobacillus acidoterrestris spores in different heating media

The aim of this work was to study the influence of temperature (85, 90, 95 and 100 °C), total soluble solids (SS: 10 and 20°Brix or % by weight of sucrose) and pH (3.5 and 4.0) on decimal reduction time ( D- value) of the Alicyclobacillus acidoterrestris strain DSM2498 spores in apple juice, orange juice and malt extract broth (MEB). The effects of SS and pH on D -values and z- values in each media were insignificant ( P  > 0.05). In apple juice, orange juice and MEB, z- values of A. acidoterrestris for pH 3.5 and pH 4.0 were 12.2 ± 1.3–14.2 ± 3.2 °C, 11.2 ± 0.3–9.4 ± 0.0 °C and 11.9 ± 0.8–10.3 ± 0.4 °C, respectively. z- values of apple juice, orange juice and MEB samples with SS = 10°Brix and SS = 20°Brix were 14.1 ± 3.2–12.2 ± 1.3 °C, 10.2 ± 0.7–10.5 ± 1.1 °C and 11.3 ± 1.5–10.9 ± 0.2 °C, respectively. However, D -values of all samples were affected by temperature significantly ( P  < 0.01). Average D -values of apple juice, orange juice and MEB were 101.2 ± 14.7, 34.4 ± 7.9, 20.3 ± 4.9 and 4.3 ± 1.3 min for 85, 90, 95 and 100 °C. This study demonstrated that A. acidoterrestris spores exhibited high resistance to thermal processing applications. pH and SS of the media did not affect thermal resistance.     

Effects of pH, cinnamaldehyde and heat-treatment time on spore viability of Alicyclobacillus acidoterrestris

A three variables–five levels central composite design was used to study the effects of pH (3.5, 4.0, 4.5, 5.0 and 5.5), cinnamaldehyde (0, 40, 80, 120 and 160 ppm) and heating temperature (80, 84, 88, 92 and 96 °C) on the spores of Alicyclobacillus acidoterrestris in malt extract broth (MEB). The heat shock resulted in a slight decrease of spore number from 3.2 to 2.5 log CFU mL⁻¹ and occurred at 80–85 °C depending on the pH of the medium. Otherwise, cinnamaldehyde acted as an additional hurdle within the storage time; the critical amounts to be used were included in the range 41.35–44.42 ppm of cinnamaldehyde and were related positively to the pH, i.e. the critical amount of active compound decreased with the decreasing of the pH.     

Modeling the combined effects of pH, temperature and ascorbic acid concentration on the heat resistance of Alicyclobacillus acidoterrestis

In this study, thermal inactivation parameters (D- and z-values) of Alicyclobacillus acidoterrestris spores in McIlvaine buffers at different pH, apple juice and apple nectar produced with and without ascorbic acid addition were determined. The effects of pH, temperature and ascorbic acid concentration on D-values of A. acidoterrestris spores were also investigated using response surface methodology. A second order polynomial equation was used to describe the relationship between pH, temperature, ascorbic acid concentration and the D-values of A. acidoterrestris spores. Temperature was the most important factor on D-values, and its effect was three times higher than those of pH. Although the statistically significant, heat resistance of A. acidoterrestris spores was not so influenced from the ascorbic acid within the concentration studied. D-values in apple juice and apple nectars were higher than those in buffers as heating medium at similar pH. The D-values ranged from 11.1 (90 °C) to 0.7 min (100 °C) in apple juice, 14.1 (90 °C) to 1.0 min (100 °C) in apple nectar produced with ascorbic acid addition, and 14.4 (90 °C) to 1.2 min (100 °C) in apple nectar produced without ascorbic acid addition. However, no significant difference in z-values was observed among spores in the juices and buffers at different pH, and it was between 8.2 and 9.2 °C. The results indicated that the spores of A. acidoterrestris may survive in fruit juices and nectars after pasteurization treatment commonly applied in the food industry.     

酸土环脂芽孢杆菌检测与控制技术研究进展

酸土环脂芽孢杆菌是造成果汁变质的一种主要腐败菌。在生产实践中快速、准确的检测手段必不可少,同时对该菌进行控制是提高产品质量、减少污染的重要环节,特别是对其耐热芽孢的抑制。通过对酸土环脂芽孢杆菌生长、代谢特性的分析,重点阐述了该菌污染的检测技术和菌种的分离、鉴定方法;同时从果汁生产环节入手探讨了控制该菌的各种方法,并指出今后应该进行的研究方向。     

脂环酸芽孢杆菌(Alicyclobacillus)分离鉴定研究进展

脂环酸芽孢杆菌(Alicyclobacillus)由于其耐热、嗜酸的独特生存能力而受到全球学术界和食品工业界的极大关注,本文系统介绍与分析了脂环酸芽孢杆菌的特征、结构、分类、分离方法、形态学、生理生化、生长条件、细胞膜脂肪酸组分分析、甲基萘醌分析、DNA组成及杂交试验、16S rRNA/DNA测序及系统发育分析鉴定方法,提出了我国应着力进行的几个研究领域:①脂环酸芽孢杆菌属新菌资源的开发;②脂环酸芽孢杆菌属细胞膜中ω-环状脂肪酸和藿烷类化合物的功能研究;③脂环酸芽孢杆菌属快速检测方法研究;④脂环酸芽孢杆菌属各菌种代谢动力学和基于代谢产物的快速检测技术;⑤脂环酸芽孢杆菌属控制技术方法研究;⑥脂肪酸芽孢杆菌热稳酶研究及工业化应用.     

Determination of guaiacol produced by Alicyclobacillus acidoterrestris in apple juice by using HPLC and spectrophotometric methods, and mathematical modeling of guaiacol production

In this study, easy detection of Alicyclobacillus acidoterrestris was performed by determination of guaiacol in apple juice. Guaiacol produced by A. acidoterrestris was determined by using HPLC, UV-Vis spectrophotometer, and Minolta spectrophotometer. Statistical analysis showed that the methods used for measuring the guaiacol concentrations were not significantly different (p > 0.05). Guaiacol formation in apple juice spiked with different levels of A. acidoterrestris spores was also analyzed using Gompertz, Logistic, and Richards models. In all cases, a good agreement between experimental data and fitted values was obtained. Using the modified Gompertz model, the derived biological parameters were calculated. Guaiacol formation rates (μ) and final guaiacol concentrations (A) were very similar in all cases, regardless of the initial A. acidoterrestris spore counts. However, lag phase durations (λ) were found to be dependent on the initial bacterial counts, and increased from 28.4 to 37.6 h, when initial inoculation level decreased from ∼10³ to ∼10¹ cfu/mL.     

脂环酸芽孢杆菌的研究进展

概述了脂环酸芽孢杆菌的特性和分类,介绍和分析了脂环酸芽孢杆菌分离鉴定的方法及其对果汁工业的危害情况及控制方法的研究进展。     

脂环酸芽孢杆菌的培养基筛选及优化

比较了脂环酸芽孢杆菌A.acidotcrrstris DSMZ 3922的芽孢在PDA、K培养基和SK培养基中的复苏情况,结果表明,SK培养基是三者中最适合于脂环酸芽孢杆菌生长的培养基;用正交试验L9(33)研究了Ca2+、Mn2+以及Tween 80在SK培养基中的添加浓度对A.acidotcrrstris DSMZ 3922、A.acidiphilus DSMZ14558生长情况的影响,结果表明,3因素对A.acidotcrrstris DSMZ 3922及A.acidiphilus DSMZ 14558生长情况影响的主次顺序均依次为Ca2+>Mn2+>Tween 80.在SK培养基中添加1.5g/L的Ca2+,0.1ppm的Mn2+,以及0.75mL/L的Tween 80是对A.acidotctrstris DSMZ 3922生长较为有利的组合;在SK培养基中添加1.5g/L的Ca2+,0.1ppm的Mn2+,以及0.25mL/L的Tween 80是对A.acidiphilus DSMZ 14558生长较为有利的组合.