耐热型植物乳杆菌微胶囊粉剂

目的:提高植物乳杆菌的耐热性。方法:在前期研究的微胶囊乳液中加入2-羟丙基-β-环糊精(2-HP-β-CD),通过喷雾干燥法制备植物乳杆菌微胶囊粉剂。通过响应面法,对不同参数进行优化。采用光学显微镜、电子显微镜对其形态进行观察,通过激光粒度分析仪及Zeta电位分析仪测定其粒径电位,利用分光光度计测定其吸光度变化,并采用烘箱加热的方式进行耐热性试验。结果:随2-HP-β-CD与乳液质量比、进料速率、进风温度、进风速率增加,存活率呈先升后降的趋势(P<0.05);在2-HP-β-CD与乳液质量比为3.13、进料速率为288 mL/h、进风速率为112 m³/h、进风温度为118℃时,菌体存活率为(99.53±0.11)%,载菌量为1.64×10⁹ CFU/g;扫描电镜和显微结果显示微胶囊为中心凹陷的微粒,在水中呈球形分散,菌体聚集于内部,其水分散液吸光值1 h内变化不显著,Z-平均粒径为472.7 nm且粒径分布较集中,Zeta电位为-40.37 mV;微胶囊在130℃处理60 s,存活率为96.79%,与菌泥差异极其显著(P<0.0...     

植物乳杆菌发酵桑葚酵素工艺优化及质量评价

目的:解决新鲜桑葚难以保存的问题,将新鲜桑葚制成桑葚酵素。方法:以新鲜桑葚汁为原料,植物乳杆菌为生产菌种,总酚含量为评价指标,通过单因素试验结合响应面试验优化了植物乳杆菌桑葚酵素的发酵工艺,并对桑葚酵素的理化、微生物及感官质量指标进行评价。结果:优化后发酵工艺条件为发酵时间40 h、发酵温度32℃、接种量25%,所制得的植物乳杆菌桑葚酵素的总酚含量达(43.48±0.67)μg/mL,是未经发酵的桑葚汁的1.62倍,可溶性固体物含量为5.36%,pH值为4.08±0.01,微生物指标满足国家标准;桑葚酵素呈紫红、色泽均匀,具有浓郁的桑葚果香和发酵的香味、无异味,酸味柔和、风味好,有光泽、无杂质及沉淀。结论:经植物乳杆菌发酵制得桑葚酵素的过程有生物活性物质产生,有利于提高桑葚酵素质量。     

基于复合诱变选育高抑菌活性植物乳杆菌

为了获得高抑菌性能的乳酸菌,以植物乳杆菌DY6为出发菌株,首先对其进行常压室温等离子体（atmospheric room temperature plasma, ARTP）和亚硝基胍（nitrosoguanidine,NTG）的复合诱变,获得了抑菌活性较强的诱变菌株AN-55、AN-58及AN-68,其抑菌活性分别提高了19.83%、21.91%和18.40%。随后,经遗传稳定性实验发现菌株AN-55的抑菌性能更加稳定,8次传代后其抑菌活性较野生型菌株仍能提高20.51%。与此同时,菌株AN-55的基因组重测序结果表明plnK、cps4G、pts9D等基因序列的改变可能对提高菌株抑菌活性有重要影响,为植物乳杆菌在天然防腐剂和抗生素替代物的应用奠定了理论基础。     

Pediocin PA-1 and a pediocin producing Lactobacillus plantarum strain do not change the HMA rat microbiota

The bacteriocin pediocin PA-1 has potential use as a food biopreservative, and understanding its effect on the commensal gut microbiota is important for assessment of consumer risks associated with the use of biopreservative cultures. Effects of ingested (i) pediocin PA-1 producing Lactobacillus plantarum DDEN 11007, (ii) the plasmid cured pediocin negative L. plantarum DDEN 12305, or (iii) supernatants of either of these two strains on the composition of the intestinal microbiota of Human Microbiota Associated (HMA) rats were examined by selective cultivation and molecular methods. The culturable microbiota was in all treatments dominated by lactic acid bacteria and coliforms and no changes in the rat commensal microbiota were detected after ingestion of either of the two L. plantarum strains as determined by both culturable methods and molecular methods (DGGE). Both strains were detected in the faeces after ingestion. Pediocin PA-1 did not mediate changes of the rat microbiota, and a biological assay indicated that the bacteriocin was degraded or inactivated during passage through the intestine.     

Stress resistance of biofilm and planktonic Lactobacillus plantarum subsp. plantarum JCM 1149

The purpose of this study was to investigate the change in resistance of biofilm and planktonic food spoilage lactic acid bacteria (LAB) to environmental stresses, which strongly inhibit bacterial growth and are important in food preservation or in disinfection. The stress responses of biofilm and planktonic cells of Lactobacillus plantarum subsp. plantarum JCM 1149, which was used as a model spoilage bacterium, in various organic acids (namely, acetic acid, citric acid, lactic acid, and malic acid), ethanol, and sodium hypochlorite, were investigated using survival tests. The bacterial cells in biofilms showed greater resistance to all treatments than the planktonic bacterial cells in either the stationary or logarithmic phase. The planktonic bacterial cells showed reduced resistance to acetic acid after the cell suspension was diluted; however, intriguingly, the bacterial cells in biofilms maintained their resistance to acetic acid even after they were suspended or the cell suspension was diluted. These findings suggested the risk for food spoilage due to LAB derived from biofilms and suspended or diluted in foods, and demonstrated the importance of controlling biofilms of LAB in the food industry.     

Effect of a single oral administration of Lactobacillus plantarum DSMZ 8862/8866 before and at the time point of weaning on intestinal microbial communities in piglets

The aim of this study was to evaluate whether a single administration of two strains of Lactobacillus plantarum (DSMZ 8862 and 8866) either before or at the time point of weaning can influence the intestinal microbiota of piglets. A total of 176 piglets were allocated into five groups: control (LP0), administration of 5 × 10⁹ or 5 × 10¹⁰ cfu at day 25 of life (LP1, LP2) and administration of 5 × 10⁹ or 5 × 10¹⁰ cfu at day 28 of life (LP3, LP4). Piglets were weaned on day 28 of life. On day 25 (LP1, LP2), 28 (LP0, LP3, LP4), 33 (all groups) and 39 (all groups) of life, 10–13 animals of each group were killed and genomic DNA was extracted from small and large intestinal contents. Denaturing gradient gel electrophoresis demonstrated that administration of L. plantarum had a significant effect in GIT microbial communities as revealed by the Simpson's index of diversity and cluster analysis based on the Dice similarity index; this effect was more pronounced in groups LP3 and LP4. A treatment dependent presence of Clostridium glycolicum-like, Lactobacillus sobrius-like, Eubacterium rectale-like and Roseburia faecalis-like phylotypes was observed. The results show that the administration of L. plantarum at the point of weaning can influence gastrointestinal microbiota in weaning piglets which may have positive results on gastrointestinal health.     

HACCP在方便面生产中的应用

本文探讨了HACCP在方便面生产中的应用,通过对生产各工序的危害分析,建立了卫生标准操作规范(SSOP)、关键控制点、关键限值、控制方法和验证程序,顾客投诉率下降40％。     

Survival of human derived Lactobacillus plantarum in fermented cereal extracts during refrigerated storage

The aim of this study was to investigate the survival of a potentially probiotic Lactobacillus plantarum strain in barley, wheat and barley malt extracts. The extracts were produced from three flour/water suspensions, i.e., 5%, 20%, 30% w/w. After inoculation, the cultures were incubated for 24 h at 37 °C, and were subsequently stored at 4 °C for up to seventy days. The lactic acid and reducing sugar concentrations at the beginning of storage were significantly different between the fermented media, ranging from 0.5 g/L to 17 g/L and from 0.8 g/L to 6.5 g/L respectively, while the pH ranged between 2.9 and 3.4. It was observed that the cells survived much better in the malt extracts compared to barley and wheat extracts during refrigerated storage. Based on the results from a study using model media and supplemented cereal extracts it was derived that this was most likely due to their higher sugar concentration and the presence of protective unidentified compounds, albeit the fact that the malt extracts contained higher amounts of lactic acid.     

不同来源的植物乳杆菌基因组和抗生素抗性

为了研究不同分离源对植物乳杆菌基因组和功能的影响,选取了来自发酵酱、泡菜、粪便3种环境的33株植物乳杆菌,通过比较基因组学手段研究菌株的基因组基本特征、直系同源基因、系统进化关系,并结合功能基因注释结果与表型结果分析菌株对抗生素的耐药性。系统发育树揭示了分离源对植物乳杆菌的遗传进化具有较为显著的影响;同源基因结果表明粪便源的菌株其特殊基因数量要高于泡菜源和发酵酱源菌株的数量。33株植物乳杆菌均含有环丙沙星、四环素、氯霉素、甲氧苄氨嘧啶和万古霉素的抗性基因,菌株对这些抗生素也表现出抗性;绝大多数菌株对庆大霉素、链霉素、卡那霉素、新霉素、氨苄西林敏感,在基因组中也没有相关抗性基因;而红霉素和克林霉素的基因和表型并不对应。抗生素实验结果说明,分离源对菌株影响较小,大部分基因型与表型可以对应,基因组学对研究植物乳杆菌的生理特性起一定的指导作用。     

植物乳杆菌微胶囊化研究

为保护植物乳杆菌的活性以增强乳杆菌在动物肠道内的益生功能,以天然发酵玉米青贮饲料中优良植物乳杆菌作为芯材,乳清蛋白和明胶为壁材,利用喷雾干燥法制成微胶囊,并以植物乳杆菌包埋率为响应值,研究壁材配比、壁材添加量、进风温度、进料量4个因素,进行中心组合实验(Box-Behnken),通过响应面分析对喷雾干燥法制备植物乳杆菌微胶囊条件进行优化。结果表明:最优条件为壁材配比(乳清蛋白与明胶质量比)1:2、壁材添加量22%、进风温度127℃、进料量35%,在此条件下,植物乳杆菌包埋率为62.15%。结论:本研究为应用喷雾干燥法制备植物乳杆菌微胶囊奠定了基础。     

植物乳杆菌发酵提取未成熟琯溪蜜柚果实果胶工艺优化

目的:利用植物乳杆菌发酵未成熟琯溪蜜柚果实提取果胶,并分析植物乳杆菌发酵提取果胶的性质。方法:以果胶提取率为指标,采用正交试验优化植物乳杆菌发酵未成熟琯溪蜜柚果实提取果胶的工艺条件,并测定果胶的半乳糖醛酸含量、酯化度、蛋白质、持水性、持油性、乳化活性以及乳化稳定性。结果:植物乳杆菌发酵未成熟琯溪蜜柚果实提取果胶的最佳工艺条件为发酵温度37℃、植物乳杆菌发酵接种量14%、液料比25∶1 (mL/g)、发酵时间12 h。此条件下未成熟琯溪蜜柚果胶提取率为11.60%;果胶的半乳糖醛酸含量、酯化度、蛋白质、持水性、持油性、乳化活性以及乳化稳定性分别为26.13%、68.58%、1.57%、0.53 g/g、7.01 g/g、14.33%和33%。结论:植物乳杆菌发酵提取的果胶提取率与酸法工艺的相似,所得果胶为高酯化度果胶,且应用性质良好。     

Functional properties of Lactobacillus plantarum strains: A multivariate screening study

Thirty-two Lactobacillus plantarum strains isolated from different sources were genetically characterized at subspecies level with recA gene based multiplex PCR and pulsed-field electrophoresis.     

Effect of microencapsulation on survival of Lactobacillus plantarum in simulated gastrointestinal conditions, refrigeration, and yogurt

In the present research the survival of free and microencapsulated cells of a new strain of Lactobacillus plantarum BL011 under stress conditions was tested in sodium alginate or pectin, coated with sodium alginate or chitosan. Results for the simulated gastrointestinal medium (SGT) showed no change in viability of cells in relation to the control. However, the simulated gastric medium (GM) drastically reduced the viability under the tested conditions, with no significant differences between free and immobilized cells. Under refrigerated storage viability of immobilized cells were greatly enhanced compared to the free microorganisms, and the treatments showing the lowest loss of viability were those of 4% (w/v) pectin, 3% (w/v) sodium alginate coated with chitosan and a mixture of 2% (w/v) sodium alginate and 2% (w/v) pectin, respectively. Loss of viability of immobilized L. plantarum in 3% alginate coated with chitosan in yogurt was of 0.55 log cycles during 38 days of storage. The results of this study suggest the efficiency of immobilization techniques to increase the survival of lactobacilli in yogurt under refrigerated storage.     

Production of -aminobutyric acid (GABA) by Lactobacillus paracasei isolated from traditional fermented foods

Lactobacillus strains that accumulated γ-aminobutyric acid (GABA) in culture medium were screened to determine strains with high GABA-producing ability. One strain, NFRI 7415, which was isolated from a Japanese traditional fermented fish (funa-sushi), showed the highest GABA-producing ability among the screened strains. Identification tests (i.e., 16S rDNA sequencing and sugar assimilation ability) indicated that NFRI 7415 belongs to Lb. paracasei. The GABA production was further improved by the addition of pyridoxal phosphate to the culture medium and pH regulation of culture medium at pH 5.0. Under optimal cultivation conditions, strain NFRI 7415 produced GABA at a concentration of 302 mm when the glutamate concentration in the culture medium was 500 mm.     

Effect of Lactobacillus plantarum and chitosan in the reduction of browning of pericarp Rambutan (Nephelium lappaceum)

The effects of Lactobacillus plantarum alone or in combination with chitosan were evaluated on quality and color retention in rambutan fruits (Nephelium lappaceum) stored at 25 °C and 10 °C with 75 ± 2.5% of relative humidity for 10 and 15 days, respectively. The development of the microorganisms was evidenced by viability analyses and lactic acid production. The application of L. plantarum significantly improved color retention (a* and L*), and reduced weight losses. The lactobacilli, alone or in combination with chitosan, preserved fruit quality characteristics such as firmness, total soluble solids and titratable acidity. The lactobacilli application on rambutan pericarp produced acidification of pericarp and avoided the browning; thereby desiccation was prevented due to biofilm formation.     

Bacteriocinogenic Lactobacillus plantarum ST16Pa isolated from papaya (Carica papaya) — From isolation to application: Characterization of a bacteriocin

Strain ST16PA, isolated from papaya was identified as Lactobacillus plantarum based on biochemical tests, PCR with species-specific primers and 16S rDNA sequencing. L. plantarum ST16PA produces a 6.5 kDa bacteriocin, active against different species from genera Enterobacter, Enterococcus, Lactobacillus, Pseudomonas, Streptococcus and Staphylococcus and different serotypes of Listeria spp. The peptide is inactivated by proteolytic enzymes, but not when treated with ??-amylase, catalase, lipase, Triton X-100, SDS, Tween 20, Tween 80, urea, NaCl and EDTA. However, presence of 1% Triton X-114 deactivates the bacteriocin. No change in activity was recorded after 2 h at pH values between 2.0 and 12.0, and after treatment at 100 °C for 120 min or 121 °C for 20 min. The mode of activity against Lactobacillus sakei ATCC 15521, Enterococcus faecalis ATCC 19443 and Listeria innocua 2030C was bactericidal, resulting in cell lysis and enzyme-leakage. No significant differences in cell growth and bacteriocin production were observed when strain ST16Pa was cultured in MRS broth at 26 °C and 30 °C for 24 h (25 600 AU/ml). However, even though strain ST16PA grows well in MRS broth at 15 °C and 37 °C, a reduction of bacteriocin production was observed (400 AU/ml and 1600 AU/ml, respectively). In addition, effect of MRS medium components, different initial pH and additions of glycerol or vitamins to the media on bacteriocin ST16Pa production was studied.Peptide ST16PA adsorbs (400 AU/ml) to producer cells. However, bacteriocin ST16Pa was adsorbed at 50% to cells of L. innocua 2030C and at 75% to L. sakei ATCC 15521 and E. faecalis ATCC 19433 when experiments were conducted at 30 °C and pH 6.5. Adsorption of bacteriocin ST16Pa to target cells at different temperatures, pH and in presence of potassium sorbate, sodium nitrate, sodium chloride, ascorbic acid, Tween 80 and Tween 20 were also studied. To the best of our knowledge, this is the first report on detection of L. plantarum in papaya.     

Inhibition by Lactobacillus sakei of other species in the flora of vacuum packaged raw meats during prolonged storage

The abilities of five Lactobacillus sakei strains and one Lactococcus lactis strain to retain inhibitory activity against several target organisms in the flora of product during 12 weeks storage of vacuum-packaged lamb and beef was investigated. L. sakei strains were generally found capable of developing dominant populations on both beef and lamb. L. lactis 75 grew poorly on lamb did not inhibit co-inoculated Brochothrix thermosphacta. Lamb inoculated with the Sakacin-A producer L. sakei Lb706 had lower Listeria monocytogenes populations than lamb inoculated with a bacteriocin-negative variant. In beef packs inoculated with Clostridium estertheticum spores and L. sakei strain 27, 44 or 63, the development of blown-pack spoilage was delayed by up to one week. Campylobacter jejuni inoculated onto beef was recovered from fewer packs when it was co-inoculated with 3000 CFU cm⁻² of L. sakei strain 27, 44 or 63. Observed inhibition did not always correlate with inhibition observed in earlier media-based studies, supporting the view that functionality identified using simple media-based screening methods may not be replicated in the complex environment of stored foods, and vice-versa. These findings further define a set of L. sakei strains with potential for the extended bio-preservation of minimally processed fresh beef and lamb.     

植物乳杆菌ZU018增殖培养基的优化

本研究以植物乳杆菌ZU018为研究对象,最终活菌数为主要参考指标,对其发酵培养基成分进行了优化。采用单因素实验选择碳源、氮源的种类及优化浓度,通过部分因子试验设计初步确定了麦芽糖、酵母浸粉及柠檬酸三铵为培养基中最主要的三个影响因素,进一步运用最陡爬坡试验及Box-Benhnken试验设计对培养基组分进行优化。结果表明,最佳优化培养基配方为:麦芽糖30.03 g/L、酵母浸粉37.50 g/L、牛肉浸粉25.00 g/L、柠檬酸三铵4.39 g/L、磷酸氢二钾2.00 g/L、乙酸钠5.00 g/L、硫酸镁 0.20 g/L、硫酸锰0.05 g/L以及1.00 g/L的吐温80。最终发酵液中植物乳杆菌ZU018活菌数相比优化前提高4.86倍,达到10.67×10⁹ CFU/mL,为后续植物乳杆菌高密度发酵及应用提供了理论依据。     

Selection of Lactobacillus plantarum strains to use as starters in fermented table olives: Oleuropeinase activity and phage sensitivity

Fermented table olives (Olea europaea L.) are largely diffused in the Mediterranean area. Olives are picked at different stages of maturity and after harvesting, processed to eliminate the characteristic bitterness caused by the presence of the oleuropein glucoside and to become suitable for human consumption. The spontaneous fermentation of table olives mainly depends on lactic acid bacteria (LAB), and in particular on Lactobacillus plantarum which plays an important role in the degradation of oleuropein. The hydrolysis of oleuropein is attributed to the β-glucosidase and esterase activities of the indigenous LAB microflora. This study investigated the potential of L. plantarum strains isolated from dairy products and olives to be used as starters for fermented table olives. Forty-nine strains were typed by RAPD-PCR and investigated for the presence of the β-glucosidase (bglH) gene. The full sequence of the bglH gene was carried out. All the 49 L. plantarum strains were also tested for phage resistance. A total of six strains were selected on the basis of genotypic polymorphism, bglH gene sequence analysis, and phage resistance profile. These strains were further characterized to assess the acidifying capability, the growth at different temperatures, the tolerance to different NaCl concentrations, and the oleuropeinolytic activity. Although further characterizations are required, especially concerning the influence on sensory properties, L. plantarum proved to have the potential to be used as a debittering and fermentative agent in starter culture for fermented table olives.     

白酒酿造过程中Lactobacillus panis乳酸合成关键环境因子的鉴定

Lactobacillus panis是酱香型白酒酿造过程中主要产乳酸微生物。然而,不同环境因子对L.panis乳酸合成及乳酸合成途径关键基因的表达尚未有相关研究。作者分析了温度、乳酸、乙醇及葡萄糖4种不同的发酵过程相关的典型环境因子对L.panis乳酸产量的影响。结果表明,不同环境条件能够显著调节L.panis的生长和乳酸产量,且乳酸和乙醇是酿造过程中其乳酸合成的关键影响因素。当添加10.0 g/L乳酸后,L.panis的乳酸产量最高,达到12.3 g/L;当添加体积分数4.0%乙醇后,L.panis的乳酸产量最低,仅为7.2 g/L。使用实时荧光定量PCR方法对不同条件下L.panis乳酸合成相关基因的表达进行分析,结果表明添加乳酸能够显著上调包括编码L-乳酸脱氢酶和D-乳酸脱氢酶的ldhL和ldhD在内的乳酸合成基因的表达。然而,添加乙醇会导致L.panis乳酸合成相关基因的整体显著下调。分析调控乳酸合成的关键环境因素,对于白酒酿造过程中乳酸和酿造微生物群落调控及白酒质量控制具有重要意义。