短乳杆菌与植物乳杆菌的发酵特性

【背景】目前对于酸菜发酵的研究主要关注点是植物乳杆菌(Lactobacillus plantarum),有关短乳杆菌(Lactobacillus brevis)在酸菜方面的研究报道很少。【目的】为了挖掘短乳杆菌的发酵性能并开发酸菜发酵剂,将2株短乳杆菌分别与1株植物乳杆菌进行组合并发酵酸菜,分析短乳杆菌对酸菜发酵品质的影响。【方法】分别测定短乳杆菌与植物乳杆菌的单菌株生长产酸性能、耐酸性及亚硝酸盐降解力,并将两菌种组合后发酵酸菜,分析1-7d内酸度、乳酸菌活菌数、亚硝酸盐含量及酸菜质构特性的变化趋势。【结果】相较于短乳杆菌Lb-9-2,短乳杆菌Lb-5-3的生长和产酸速率较慢、酸耐受力较弱,但其亚硝酸盐降解力较强。两株短乳杆菌分别与植物乳杆菌Lp-9-1组合后产酸力显著增强,并在3 d时达到最低pH值(约3.10);植物乳杆菌Lp-9-1的添加使酸菜中总体乳酸菌生长延迟,在5 d时达到最高活菌数;组合菌种的样品中亚硝酸盐含量在1-7 d内变化较为平缓,前5天内两个组合之间差异不显著;接种乳酸菌会降低酸菜硬度和弹性,发酵3d时Lb-5-3/Lp-9-1组合的硬度最大,感官评价得分最高。【...     

温度和渗透应激对植物乳杆菌稳定性的影响

在植物乳杆菌对数期后期,高温（43—47℃）、低温（15—25℃）及10-30g/L NaCl等应激处理60min均可使植物乳杆菌耐热性和耐酸性得到较大的提高。其中,尤以高温应激45℃应激处理效果最好,细胞耐热残存率和耐酸残存率较对照分别提高124％和56．8％。     

植物乳杆菌对2型糖尿病大鼠的影响及影响机制

探究植物乳杆菌对2型糖尿病大鼠的影响及影响机制。对雄性SD大鼠采用高脂饮食和腹腔注射链脲佐菌素(STZ)的方法诱导其建立2型糖尿病动物模型,将建模成功的大鼠随机分为糖尿病组、高剂量干预组(1.016×10⁸CFU/100 g·BW)和低剂量干预组(2.4×10⁷CFU/100 g·BW),将由普通饲料喂养的大鼠作为对照组。干预过程中对大鼠的体重、空腹血糖(FBG)、餐后两小时血糖(2hPBG)以及糖耐量等数据进行测定。第15周后检测血脂和血清胰岛素(INS)水平,评估胰岛素抵抗指数(HOMA-IR),对大鼠粪便进行高通量测序,利用Western Blot方法测定糖脂代谢通路及胰岛素信号传导通路中相关蛋白的表达。植物乳杆菌显著提高了糖尿病大鼠体内的糖耐量、胰岛素耐量水平和HOMA-IR,改善了血脂水平异常的情况;增加了大鼠肠道菌群的多样性及丰富度,优化了大鼠肠道菌群的结构及组成,使有害菌减少,有益菌增多;显著上调PI3K、AKT蛋白表达;下调PCK1、GSK-3β蛋白表达;显著上调P-AMPK、P-ACC1蛋白表达,下调SREBP1蛋白表达,其...     

昂立植物乳杆菌及其抑菌物质的特性研究

该文研究了昂立植物乳杆菌(LP-Onlly)菌体及其代谢产物的抑菌性能,并对其代谢产物中的抑菌物质进行了部分理化特性的考察,发现LP-Onlly菌体对部分肠道有害菌有抑制作用,代谢产物中的抑菌物质对常见的肠道致病菌和食品腐败微生物具有广谱抑菌作用,对嗜酸乳杆菌及双歧杆菌等益生菌无抑制作用.该物质具有热稳定性,但抑菌活性受pH值的影响较大.     

植物乳杆菌高活力保存方法的研究及其应用

本研究旨在探讨植物乳杆菌的高活力保存方法,为植物乳杆菌饲料添加剂规模化、工业化生产奠定基础。采用4℃低温保存法、36℃烘干后常温保存法、阳离子活性载体保存法等3种方法对植物乳杆菌进行活力保存比较试验。以保存后活菌数不低于原值50%为参照标准,结果显示,对照组可保存15 d,4℃低温保存法可保存30 d,36℃烘干后常温保存法只能保存一周,而阳离子活性载体保存法则可保存2个月以上。结果表明,阳离子活性载体保存法可应用于植物乳杆菌饲料添加剂的规模化、工业化生产实践中。     

植物乳杆菌多孔淀粉直接包埋工艺的研究

利用多孔淀粉的吸附特性,将植物乳杆菌包埋于多孔淀粉内,通过测定多孔淀粉对植物乳杆菌的包埋率,研究菌体浓度、多孔淀粉添加量、振荡转速、包埋温度、pH值、时间对包埋率的影响,确定最适包埋条件。结果表明:菌体浓度10~8cfu/mL,多孔淀粉添加量为2%,pH 6.0、20℃,200 r/min振荡处理40 min,在此条件下包埋率为79.5%,对包埋后的菌体进行喷雾干燥试验,其存活率较未包埋的从0.35%提高到29.5%。     

羊骨酶解液的植物乳杆菌发酵

为进一步提高羊骨酶解液的营养价值和生物利用度,探讨乳酸菌发酵对钙离子释放、短肽形成以及酶解液抗氧化活性的影响。首先从7株乳酸菌中筛选出了产蛋白酶能力最强的植物乳杆菌,接种羊骨酶解液,以发酵液中Ca~(2+)浓度为指标,响应面法优化得到发酵工艺:酶解液中添加1%的麦芽糖,调节起始pH为5.5,以4%接种量接种驯化好的植物乳杆菌,37℃摇床发酵14 h。发酵过程中,植物乳杆菌活菌数与Ca~(2+)含量(R=0.495,P0.01)和短肽得率(R=0.655,P0.01)呈极显著正相关,而多肽生成量与短肽得率呈负相关(R=–0.013)。发酵液中的Ca~(2+)浓度(P0.05)、水解度(P0.01)、短肽得率(P0.01)、羟脯氨酸含量(P0.01)均显著高于酶解液(P0.01),植物乳杆菌活菌数达到94.6×10~8 CFU/m L。发酵还可使酶解液对DPPH、·OH、O_2~-·三种自由基的清除能力显著提高(P0.01,P0.05)。综上所述,以植物乳杆菌发酵羊骨酶解液可进一步促进骨钙的转化和胶原短肽的生成并显著提高其体外抗氧化能力,短肽和Ca~(2+)反过来促进植物乳杆菌的繁殖,增强了酶解液的益生功能,乳酸菌产生的维生素、胞外多糖等物质使羊骨酶解液更富营养。     

昂立植物乳杆菌对腹腔感染大鼠肠道微生态的影响

目的探讨经肠道补充昂立植物乳杆菌(LP-Onlly)对腹腔感染大鼠肠道微生态的影响.方法大鼠经盲肠结扎穿孔法及颈静脉空肠置管制成腹腔感染模型后,分别给予肠外营养(PN组)和PN 昂立植物乳杆菌(LP-Onlly组)持续5 d,第6天处死,取盲肠内粪便进行肠菌群培养计数及细菌种群DNA指纹图谱分析.结果 LP-Onlly组大鼠肠道内的乳杆菌和双歧杆菌数量较单纯PN组的大鼠为多,而肠道内的潜在致病菌产气荚膜梭菌数量较单纯PN组的大鼠少,DNA指纹图谱也显示LP-Onlly组大鼠优势菌群的基因条带和正常大鼠具有较高的一致性,而PN组则差异有显著性.结论 LP-Onlly能纠正腹腔感染大鼠PN时的肠道菌群紊乱, 调理肠道微生态平衡.     

短乳杆菌葡萄糖异构酶固定化的研究

采用离子交换树脂ＤＥＡＥ纤维素对短乳杆菌葡萄糖异构酶进行固定化,并用固定化细胞使葡萄糖异构化为果糖,研究了制备固定化细胞最佳条件,而且与戊二醛交联法,海藻酸钙包埋法作了比较。还研究了固定化细胞的性质及连续转化的最佳条件,用ＤＥＡＥ纤维素固定化具有酶活力高,回收率高,机械强度好,成本低等优点。     

Synthetic reconstruction of extreme high hydrostatic pressure resistance in Escherichia coli

Although high hydrostatic pressure (HHP) is an interesting parameter to be applied in bioprocessing, its potential is currently limited by the lack of bacterial chassis capable of surviving and maintaining homeostasis under pressure. While several efforts have been made to genetically engineer microorganisms able to grow at sublethal pressures, there is little information for designing backgrounds that survive more extreme pressures. In this investigation, we analyzed the genome of an extreme HHP-resistant mutant of E. coli MG1655 (designated as DVL1), from which we identified four mutations (in the cra, cyaA, aceA and rpoD loci) causally linked to increased HHP resistance. Analysing the functional effect of these mutations we found that the coupled effect of downregulation of cAMP/CRP, Cra and the glyoxylate shunt activity, together with the upregulation of RpoH and RpoS activity, could mechanistically explain the increased HHP resistance of the mutant. Using combinations of three mutations, we could synthetically engineer E. coli strains able to comfortably survive pressures of 600–800 MPa, which could serve as genetic backgrounds for HHP-based biotechnological applications.     

Effect of high hydrostatic pressure on the porin OmpC from Escherichia coli

Porin OmpC from Escherichia coli was reconstituted in liposomes and its gating kinetics were recorded at high hydrostatic pressure, up to 90 MPa, using a development of the patch clamp technique. The composition of the recording solution influenced the results but generally high hydrostatic pressure favoured channel opening.     

Examination of Lactobacillus plantarum lactate metabolism side effects in relation to the modulation of aeration parameters

AIMS: The characterization of global aerobic metabolism of Lactobacillus plantarum LP652 under different aeration levels, in order to optimize acetate production kinetics and to suppress H2O2 toxicity. METHODS AND RESULTS: Cultures of L. plantarum were grown on different aeration conditions. After sugar exhaustion and in the presence of oxygen, lactate was converted to acetate, H2O2 and carbon dioxide with concomitant ATP production. Physiological assays were performed at selected intervals in order to assess enzyme activity and vitality of the strain during lactic acid conversion. The maximal aerated condition led to fast lactate-to-acetate conversion kinetics between 8 and 12 h, but H2O2 immediately accumulated, thus affecting cell metabolism. Pyruvate oxidase activity was highly enhanced by oxygen tension and was responsible for H2O2 production after 12 h of culture, whereas lactate oxidase and NADH-dependent lactate dehydrogenase activities were not correlated to metabolite production. Limited NADH oxidase (NOX) and NADH peroxidase (NPR) activities were probably responsible for toxic H2O2 levels in over-aerated cultures. CONCLUSION: Modulating initial airflow led to the maximal specific activity of NOX and NPR observed after 24 h of culture, thus promoting H2O2 destruction and strain vitality at the end of the process. SIGNIFICANCE AND IMPACT OF THE STUDY: Optimal aeration conditions were determined to minimize H2O2 concentration level during growth on lactate.     

A study on growth characteristics and nutrient consumption of Lactobacillus plantarum in A-stat culture

Lactobacillus plantarum was grown in complex media containing glucose and yeast extract. The maximum growth yield based on yeast extract consumption was 0.5 g dwt g^-1. Growth yield Y_ATP 15–17 g dwt mol ATP^-1 was almost constant in the glucose limited A-stat experiment whereas in the yeast extract limited culture it increased with dilution rate. The maximum specific growth rate observed, 0.5 h^-1, was similar for both A-stat and batch cultures. Specific oxygen consumption, Q_O2, reached the value of 1.8 mmol O₂ h^-1 g dwt^-1. It was shown that Val, Ile, Leu, Tyr and Phe, were consumed mainly as free amino acids, while Asp, Pro, Lys and Arg were derived from peptides. Significantly more Asp, Ser, Glu, Val, Ile, Leu and Phe were consumed than needed to build up cell protein whereas some Pro, Gly, Ala and Lys was synthesized. A network of metabolic reactions in L. plantarum was proposed on the basis of the experimental data.     

Refolding of endostatin from inclusion bodies using high hydrostatic pressure

High hydrostatic pressure was used for concomitant solubilization and refolding of insoluble endostatin (ES) aggregated as inclusion bodies (IBs). High hydrostatic pressure (200 MPa or 2 kbar) was applied in combination with nondenaturing concentrations of guanidine hydrochloride. High levels of correctly folded ES (90 mg/L culture) were obtained after optimization/standardization of the procedure by applying pressures of 200 MPa for 16 h in 1.5 M guanidine hydrochloride/0.5 mM oxidized glutathione and reduced glutathione. Refolded ES was purified by affinity chromatography on a heparin column and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, size exclusion HPLC, circular dichroism, and intrinsic fluorescence. We demonstrated that high pressure can successfully convert insoluble IBs of ES expressed in Escherichia coli into an ES preparation with native tertiary structure and full biological activity.     

1株植物乳杆菌生物学特性的研究

研究1株植物乳杆菌(N3)的生物学特性,实验结果表明该菌能耐受80~85℃的高温和0.20kg/cm2蒸汽压力;直接分解玉米淀粉的乳酸产率为7.05%(36 h)和8.19%(48 h);能耐受pH为4.5的酸性环境;在人工胃液中的活菌数为4.1×106CFU/g;对金霉素、土霉素、痢特灵和氟哌酸等抗生素的敏感性强,而对防霉剂和脱霉素不敏感。植物乳杆菌(N3)是1株优良的益生素生产菌。     

The Effect of Lactobacillus plantarum Extracellular Vesicles from Korean Women in Their 20s on Skin Aging

Extracellular vesicles, which are highly conserved in most cells, contain biologically active substances. The vesicles and substances interact with cells and impact physiological mechanisms. The skin is the most external organ and is in direct contact with the external environment. Photoaging and skin damage are caused by extrinsic factors. The formation of wrinkles is a major indicator of skin aging and is caused by a decrease in collagen and hyaluronic acid. MMP-1 expression is also increased. Due to accruing damage, skin aging reduces the ability of the skin barrier, thereby lowering the skin’s ability to contain water and increasing the amount of water loss. L. plantarum suppresses various harmful bacteria by secreting an antimicrobial substance. L. plantarum is also found in the skin, and research on the interactions between the bacteria and the skin is in progress. Although several studies have investigated L. plantarum, there are only a limited number of studies on extracellular vesicles (EV) derived from L. plantarum, especially in relation to skin aging. Herein, we isolated EVs that were secreted from L. plantarum of women in their 20s (LpEVs). We then investigated the effect of LpEVs on skin aging in CCD986sk. We showed that LpEVs modulated the mRNA expression of ECM related genes in vitro. Furthermore, LpEVs suppressed wrinkle formation and pigmentation in clinical trials. These results demonstrated that LpEVs have a great effect on skin aging by regulating ECM related genes. In addition, our study offers important evidence on the depigmentation effect of LpEVs.     

Effect of temperature on gelatinization and retrogradation in high hydrostatic pressure treatment of potato starch-water mixtures

The effect of temperature (20-70 °C) on the gelatinization and retrogradation of potato starch-water mixtures (10-70%, w/w) treated with high hydrostatic pressure (HHP) (400-1000 MPa) was investigated. Gelatinization enthalpy change (ΔH_gel) and re-gelatinization enthalpy change of retrograded crystalline part (ΔH_retro) of the HHP-treated starch were evaluated using differential scanning calorimetry. The value of ΔH_gel of 10-20% (w/w) mixtures decreased with increased pressure and temperature, while ΔH_gel of 30-50% (w/w) mixtures decreased to certain values with increased pressure and the values depended on treatment temperature. With higher temperature and pressure conditions, ΔH_gel of 10-40% (w/w) mixtures reached zero, but ΔH_gel of 50-70% (w/w) mixtures did not. Retrogradation was observed with HHP-treated 20-60% (w/w) mixtures and the value of ΔH_retro depended on the starch content, pressure, and temperature. The value of ΔH_retro trended to increase with increase in starch content. In addition, retrogradation was promoted by HHP treatment at low temperature. Gelatinizaiton and retrogradation behaviors of HHP-treated (400-1000 MPa) potato starch-water mixtures (10-70%, w/w) at 20-70 °C were summerized in a series of state diagrams.     

State diagram of potato starch–water mixtures treated with high hydrostatic pressure

Potato starch–water mixture was treated with high hydrostatic pressure (HHP) of up to 1.2 GPa, and effect of starch content (10–70% (w/w)) on HHP-gelatinization was investigated by differential scanning calorimetry (DSC). Depending on the treatment pressure and potato starch content, DSC thermograms showed decrease in enthalpy change of heat gelatinization reflecting the progress of HHP-gelatinization and increase in enthalpy change of re-gelatinization of retrograded starch. From the viewpoint of the enthalpy changes, physically modified state of HHP-treated potato starch–water mixtures was classified as follows: no change, partial gelatinization, complete gelatinization, partial gelatinization and retrogradation, and complete gelatinization and retrogradation. A state diagram of potato starch–water mixtures (treatment pressure vs. starch content) was presented.