Assessment of technological attributes of autochthonous starter cultures in Turkish dry fermented sausage (sucuk)

The study aims to evaluate the technological properties of autochthonous strains (Lactobacillus sakei S15, Lactobacillus plantarum S24, L. plantarum S91, Pediococcus pentosaceus S128b and Staphylococcus carnosus G109) in Turkish dry fermented sausage (sucuk). After 24 h of fermentaPtion, all lactic acid bacteria strains reduced the pH to below 5.0, while the pH in the control group was above 5.3. The number of lactic acid bacteria strains reached 10⁸–10⁹ cfu g⁻¹ during fermentation. Staphylococcus carnosus G109 remained at the inoculation level of 10⁶ cfu g⁻¹ during ripening. Lactobacillus sakei S15 as mono-culture showed higher TBARS values compared to other strains. The control group had the lowest L* value and autochthonous strains caused no significant difference for a* value. According to principal component analysis results, most volatile compounds were positively correlated with the group containing only L. sakei S15.     

Lactobacillus plantarum BC299 can alleviate dextran sulphate sodium-induced colitis by regulating immune response and modulating gut microbiota

It is well known that probiotics are effective in relieving inflammatory bowel disease (IBD). However, their exact mechanism is still unclear. Herein, we evaluated the effect of novel Lactobacillus plantarum BC299 on dextran sodium sulphate (DSS)-induced colitis model mice and investigated the molecular mechanism. We found BC299 could regulate immune response and reduce immunosuppression by enhancing the expression of Th1 T cells in cyclophosphamide-induced immunosuppressive model mice. Meanwhile, DSS-induced colitis model mice were furtherly constructed. The results displayed BC299 can restore bodyweight and relieve colon morphology of colitis mice. Real-time quantitative polymerase chain reaction results demonstrated BC299 can regulate immune response by reducing the expression of TNF-α and IL-1β mRNA and increasing the expression of IL-10 and TGF-β mRNA. Moreover, BC299 administration reshaped and increased the diversity of gut microbiota in IBD mice. Therefore, BC299 can alleviate colitis by regulating immune response and modulating gut microbiota.     

The peptides in oat and malt extracts that are preferentially absorbed by Lactobacillus plantarum and stimulates its proliferation in milk

Lactobacillus plantarum proliferates inefficiently in milk, mainly because of its lack of cell envelope proteases and its inability to hydrolyse proteins in milk. Our previous study showed that this strain could grow well in milk with the addition of oat and malt extracts. To investigate the usage and preference for polypeptides and oligopeptides for this strain, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), o-phthaldialdehyde (OPA), high-performance liquid chromatography (HPLC), plate counting and other methods were used in this study. The results showed that proteins in fermented milk cannot be absorbed and utilised by L. plantarum, whereas polypeptides and oligopeptides provide available nitrogen sources for their growth. Short-chain peptides were more conducive to absorption and utilisation than long-chain peptides. In particular, peptides with molecular weights in the range of 200–1400 Da in the oat extract and 100–700 Da in the malt extract were preferentially absorbed and utilised.     

Dihydrofolate Reductase Thermosensitive (ts-dfrA) Mutant Induces Dihydrofolate Overproduction by Lactobacillus plantarum

Dihydrofolate (dihydrofolic acid, vitamin B9) is one of the principle reduced forms of folates, which is converted to tetrahydrofolate by dihydrofolate reductase (DHFR). The aim of the present study was to overproduce total folate by inducing accumulation of dihydrofolate (DHF) due to inhibition of DHFR in a thermosensitive mutant (TS) of Lactobacillus plantarum. This mutant (TS34) was obtained by UV-mutagenesis of L. plantarum wild-type parent strain (HMA07) isolated from fermented sausage. The mutant TS34 was characterized by its ability to grow at 30°C but not at 40°C. Strain HMA07 and its mutant TS34 were identified by determining partial sequence of 16S rRNA gene. A single C→T exchange in the dfrA (dihydrofolate reductase gene) at nucleotide 184 was detected. This exchange led to the conversion of leucine (L) to phenylalanine (F) at the position 62 in amino acid sequence (L62F transition). TS43 showed significant (p < 0.05) reduction in DHFR activity from 1.51 to 0.06 Unit/mg protein, resulting in increasing DHF amount at the expense of THF. Accordingly, total folate production by the mutant TS34 (in two-stage fermentation process with temperature shift-up from 30°C to 40°C) increased by three-fold compared with the parental strain HMA07. In conclusion, thermosensitive mutation in dfrA caused inhibition of DHFR and resulted in accumulation of DHF. The technique described here could be considered as novel strategy for overproduction of DHF in two-stage fermentation process.     

Lactobacillus Gasseri LGZ 1029 in yogurt: rheological behaviour and volatile compound composition

Yogurt preserves and enhances nutritional value of milk. In this study, we have compared several strains to determine the physicochemical, sensory, rheological and aroma characteristics of different yogurts. We used Lactobacillus gasseri LGZ 1029 (LG), commercial probiotic L. rhamnosus (LGG) and traditional fermentation strains Streptococcus thermophilus and L. bulgaricus (SL). Results showed that the flavour and texture characteristics of mixed-strain yogurts were obviously better than in single-strain yogurts. Addition of LG increased pseudoplastic behaviour, as shown by Herschel–Bulkley model analysis of rheological behaviour. The LG + SL group also had both the highest viscosity consistency index and thickening ability. In addition, a total of 57 volatile compounds were detected in yogurts and the fermentation with the addition of LG was mainly affected by ketones. Our study suggested that a yogurt with new attributes can be produced by using LGZ 1029.     

豆腐乳中乳酸菌的分离鉴定与发酵特性研究

目的分离鉴定农家自制传统发酵豆腐乳中的乳酸菌,探讨其作为豆类乳酸菌发酵饮品菌株的可行性。方法利用MRS(man rogosa sharp)培养基分离豆腐乳中的乳酸菌,通过形态学观察、生理生化特性和16S rDNA基因序列分析进行鉴定,并将所分离到的乳酸菌与适合于豆类植物发酵的植物乳杆菌FJAT-7926(Lactobacillus plantarum FJAT-7926)和干酪乳杆菌FJAT-7928 (Lactobacillus casei FJAT-7928)进行发酵特性的对比研究。结果从豆腐乳中分离出1株乳酸菌,命名为FJAT-46777,该菌株菌体细胞为圆端直杆状或圆端弯曲杆状,无芽孢,革兰氏阳性,过氧化氢酶阴性,生理生化特征与发酵乳杆菌(Lactobacillus fermentum)一致,对菌株的16S rDNA基因进行扩增测序、分子系统发育树分析,也表明其为发酵乳杆菌。发酵乳杆菌FJAT-46777发酵的豆乳中乳酸菌增殖速度最快,最终活菌数最高,为9.42 lg(CFU/mL); pH值下降最快,最终pH值最低,为3.91;滴定酸度上升速度最快,最终滴定酸度最高,为62~ΟT。结论分离自豆类自然发酵食品中的发酵乳杆菌FJAT-46777,对豆类植物为主的基质具有更优良的发酵特性,发酵时间快,乳酸菌含量高,产酸能力强,适用于发酵豆乳的开发。     

一株具降胆固醇功能的海洋源植物乳杆菌的筛选及其益生性能分析

本文采用邻苯二甲醛法从喙鲸内脏中筛选分离出一株高效降胆固醇能力的乳酸菌HJ-S2,考察并研究了乳酸菌HJ-S2的体外益生性能。通过耐酸、耐胆盐测定菌株的益生作用,研究了其对致病菌的抑菌作用及对人结肠癌细胞HT-29的粘附能力,利用形态学、API50CH和16S rDNA对菌株进行鉴定。乳酸菌HJ-S2体外降胆固醇能力达到48.82%,在p H值3的酸性环境中培养3h耐受率达到82.73%,在含0.3%的胆盐环境中培养3 h耐受率达80.62%。菌株代谢物对致病菌大肠杆菌、金黄色葡萄球菌和沙门氏菌均有一定的抑菌作用,且菌株对人结肠癌细胞HT-29很强的粘附能力,粘附细菌数为132.52,经鉴定为植物乳杆菌Lactobacillus plantarum,并命名为HJ-S2(CGMCCNO:17720)。筛选得到的植物乳杆菌HJ-S2体外具有高效降胆固醇能力,可进一步开发为功能性微生态制剂。     

副干酪乳杆菌VL8胞外多糖对巨噬细胞RAW264.7免疫激活作用研究

旨在研究副干酪乳杆菌VL8胞外多糖(Lactobacillus paracasei VL8 exopolysaccharides,VL8-EPS)对巨噬细胞RAW264.7的免疫激活作用。试验采用不同浓度的VL8-EPS作用于RAW264.7细胞,设空白对照组,脂多糖(lipopolysaccharide,LPS)处理组(终浓度为10μg/mL)和VL8-EPS处理组(终浓度为50、100、200、400、800μg/mL)。迪夫快速染色法观察细胞形态;比色法测定诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)活力;WST-1法测定超氧化物歧化酶(superoxide dismutase,SOD)活力;微量法检测超氧阴离子(O2-)的含量;钼酸显色法检测过氧化氢(H2O2)的分泌量。结果表明,VL8-EPS能够改变细胞形态,剂量依赖性提高RAW264.7细胞内iNOS、SOD酶活力,促进细胞分泌O2^-和H2O2。在800μg/mL时,较空白对照组分别提高了104.64%、10.30%、666.87%、81.78%。说明VL8-EPS通过提高胞内酶活性及释放活性氧激活RAW264.7细胞从而发挥免疫增强作用。     

1株水开菲尔粒中植物乳杆菌的鉴定及产细菌素基因分析

目的:对从水开菲尔粒中分离得到1株能产生抑菌物质的菌株QF01进行菌种鉴定,并对其产细菌素进行实验、鉴定和基因序列分析。方法:通过牛津杯法测定抑菌能力,采用聚合酶链式反应(polymerase chain reaction,PCR)扩增16S r DNA和细菌素相关基因并测序,利用Pro Param tool、TMHMM 2.0、Inter Pro Scan、SOPM和SWISSMODEL在线软件对基因编码产物进行分析。结果:该菌株产的细菌素对大肠杆菌(Escherichia coli JM109)有明显的抑制作用,通过16S r DNA序列分析初步确定该菌株属于植物乳杆菌(Lactobacillus plantarum)。该菌株含有pln D、pln EF、pln V、pln R四种基因,其中pln V基因编码产物有跨膜螺旋结构,其结构存在信号肽特征。此外,从三级结构的模型预测得到4种基因的编码产物均存在α-螺旋、β-转角、伸展链和无规则卷曲。结论:从水开菲尔粒分离得到的L.plantarum QF01菌株,含有pln D、pln EF、pln V、pln R细菌素基因,对大肠杆菌有很好的抑制作用。