降胆固醇乳酸菌的筛选及其在内蒙古发酵香肠中的应用

通过对乳酸菌降胆固醇生物学特性及对发酵肉制品中胆固醇降解作用进行研究,结果表明,从内蒙古传统肉肠中分离筛选得到的8 株乳酸菌中,菌株X3-2B有较强的胆固醇降解能力。且在MRS培养基中添加3 g/L胆盐、20 g/L胆固醇和 20 g/L葡萄糖时菌株X3-2B的胆固醇降解能力最大,在不同培养基中发酵不同时间菌株X3-2B对胆固醇的降解能力显著高于标准菌株。在以菌株X3-2B为发酵剂的发酵香肠中,其胆固醇含量显著低于对照组。故菌株X3-2B可作为一株降胆固醇性能较好的肉制品发酵剂。     

降胆固醇乳酸菌的筛选及其在内蒙古发酵香肠中的应用（英文）

通过对乳酸菌降胆固醇生物学特性及对发酵肉制品中胆固醇降解作用进行研究,结果表明,从内蒙古传统肉肠中分离筛选得到的8株乳酸菌中,菌株X3-2B有较强的胆固醇降解能力。且在MRS培养基中添加3 g/L胆盐、20 g/L胆固醇和20 g/L葡萄糖时菌株X3-2B的胆固醇降解能力最大,在不同培养基中发酵不同时间菌株X3-2B对胆固醇的降解能力显著高于标准菌株。在以菌株X3-2B为发酵剂的发酵香肠中,其胆固醇含量显著低于对照组。故菌株X3-2B可作为一株降胆固醇性能较好的肉制品发酵剂。     

天然乳酸菌对肉中胆固醇降解作用及其生长特性

通过测定从内蒙古传统乳制品中分离的6株降胆固醇性能优良的乳酸菌的生长特性及其在模拟肉汤中胆固醇降解作用,研究其在发酵肉制品中的降胆固醇性能,结果表明,菌株2-2B33,TE7401,TE5301,TF5201胆固醇降解率高达70%以上且优于标准菌。各菌株在MRS、模拟肉汤、MRS+6%NaCl+150 mg/kg NaNO2培养基中的产酸性能较好且在2 d后pH可降到3.90左右。其中TE7401号菌对胆固醇的降解率、不同温度不同pH下的生长性能及产酸性能较好,故可做为1株降胆固醇性能、生长性能良好的发酵剂。     

发酵香肠发酵剂菌种筛选的研究

从近年发酵肉制品中涉及到的有益微生物中，筛选出能够快速发酵并使发酵肠风味品质俱佳、质量稳定的发酵剂。并且通过正交试验确定发酵香肠组合发酵剂的最优配比，对香肠成熟过程和贮藏过程中的pH值、乳酸菌数量的变化、感官品质等方面进行分析，研究结果表明，植物乳杆菌（Lp）、嗜酸乳杆菌（La）和戊糖片球茼（Pp）是良好的发酵肉制品发酵剂；它们作为发酵香肠发酵剂的最优配比是0.6％，0.6％，0.3％；此组合发酵剂发酵的香肠的pH值，乳酸菌数，感官特征等指标均优于单一菌种发酵的香肠.     

肉制品发酵剂的筛选与发酵特性的研究

以MRS培养基为基础培养基,从自然发酵香肠和自制腌肉中分离、筛选出三株乳酸菌(F1、F2、F3),对其生化特性、发酵特性、生长状况、产酸能力、致死温度及菌株间的拮抗性进行研究,结果表明:3株乳酸菌均符合肉制品发酵剂的要求,可以作为肉制品发酵剂;菌株F1和F3,F2和F3可以作为肉制品的混合发酵剂.     

酸菜中降解亚硝酸盐乳酸菌的筛选、鉴定及其在发酵香肠中的应用

目的:依据多项评价指标筛选亚硝酸盐降解能力强且适用于肉制品发酵的乳酸菌。方法:首先采用添加0.3%Ca CO3的MRS培养基从15份不同产地的农家自制酸菜中筛选乳酸菌,其后以降解亚硝酸盐能力和肉制品发酵剂所需要求为筛选指标对初筛所得乳酸菌进行复筛,并对其进行菌种的分子鉴定。最后以筛选到的菌株为发酵剂制作单菌株发酵香肠,通过检测香肠在发酵和贮藏期间的Na NO2含量来进一步验证所得菌株的实际降解亚硝酸盐能力。结果:筛选到1株降解亚硝酸盐能力优良(降解率为78.77%)且适于肉制品发酵的乳酸菌菌株,16S r DNA全序列分析鉴定为鼠李糖乳杆菌(Lactobacillus rhamnosus GG,ATCC 53103)。香肠发酵实验结果显示,在发酵结束时香肠的Na NO2含量为12.2 mg/kg,低于国家标准。同时,微生物指标检测结果证明了此条件下生产的发酵香肠的质量是安全可靠的。结论:鼠李糖乳杆菌能有效降低发酵香肠中的Na NO2含量,可用于开发成用于生产低亚硝酸盐的健康、安全的肉品发酵剂。     

侗族传统酸鱼中降亚硝酸盐乳酸菌的筛选、鉴定及其在发酵香肠中的应用

以侗族传统酸鱼作为乳酸菌筛选源,分离出17株乳酸菌。以降亚硝酸盐能力为筛选指标对初筛所得乳酸菌进行复筛,筛选到2株降亚硝酸盐能力优良且适于肉制品发酵的乳酸菌菌株,经16S r DNA全序列分析鉴定2株菌分别为植物乳杆菌(DLY-7)和发酵乳杆菌(DLY-10)。以筛选到的菌株为发酵剂制作单菌株发酵香肠,结果显示,在发酵结束时发酵香肠的NaNO2含量为9.81 mg/kg(DLY-7)和11.23 mg/kg(DLY-10),均低于国家标准,表明筛选菌株可用于开发成用于生产低亚硝酸盐的健康肉制品发酵剂。     

发酵香肠益生型乳酸菌发酵剂的筛选

从主要自然发酵肉制品中分离、纯化得到77株耐酸耐胆汁的乳酸菌,并进一步对分离菌株进行了耐高酸(pH2.5)、0.3%胆盐的生长实验.25株在pH值2.5,培养3 h后表现出较高存活性,21株经低酸预处理后在0.3%胆盐存在的情况下生长良好.最后运用API 50CHL对筛选菌株进行了鉴定.这些菌株可作为进一步筛选发酵香肠优良乳酸菌种的出发菌株,以便筛选品质优良的益生性乳酸菌,作为新型发酵香肠发酵剂.     

传统发酵肉制品中降解生物胺菌株的筛选鉴定与应用研究

该研究利用高效液相色谱（HPLC）法对65株来源于传统发酵肉制品的耐盐、耐亚硝酸盐乳酸菌所产生物胺进行定性定量检测,筛选出降解率最高的不产生物胺菌株PL-ZL001。经形态观察、生理生化试验研究,并结合16S rDNA序列分析对其进行鉴定,同时探索其作为发酵剂对发酵香肠中生物胺含量的影响。结果表明,菌株PL-ZL001被鉴定为植物乳杆菌（Lactobacillus plantarum）,添加菌株PL-ZL001可抑制发酵香肠中6种生物胺的积累,尤其是对毒性最大的组胺含量的控制,效果显著优于商业用木糖葡糖球菌（Staphyloccus xylose）（P＜0.05）。     

分离自传统自然发酵食品中降胆固醇乳酸菌的筛选与评价

为了筛选优良的降胆固醇乳酸菌,本分析了从传统自然发酵食品中分离的30株乳酸菌,从中筛选出WS1、LP2、LP3、LP4、LP5和LP6 6株具有较强降胆固醇能力的乳酸菌,并通过对其胆盐耐受性及在含胆盐MRS培养基中的降胆固醇效率的分析,复筛出其中的WS1、LP2、LP3和LP6 4株乳酸菌。对4株乳酸菌的生长特征、产酸特征、不同生长阶段的种子液对胆固醇的降低效率的影响、耐酸特征及人工胃酸的耐受性进行分析和评价。结果显示,4株优选的乳酸菌降胆固醇能力较好,在胆盐中具有一定的存活力。胆盐的存在对降胆固醇能力具有一定的促进作用,4株菌在常规MRS培养基中生长和产酸特征稳定。不同生长时期的种子液对乳酸菌的降胆固醇能力有一定影响,对数期优于稳定期。耐酸性分析表明, pH越高,乳酸菌耐受性越好。pH 2.5,处理2h,乳酸菌依然具有一定的存活力;pH 3.5,乳酸菌存活力和存活时间都有所提高,pH 4.5时的存活力基本和自然pH的保持一致。在人工胃酸条件下,4株复筛乳酸菌在pH 2的条件下,存活时间超过4 h,随着pH的升高,存活率和存活时间都有所提高。本试验结果表明,4株乳酸菌均具有降胆固醇的特征,为后续优良菌株的降胆固醇机理研究打下了基础。     

3 株乳酸菌及其组合对发酵牦牛肉灌肠中肽变化的影响

本实验探讨了3 株乳酸菌发酵剂及其组合在发酵牦牛肉灌肠过程中对肽变化规律的影响。人工接种米酒乳杆菌（Lactobacillus sake,L. s）、植物乳杆菌（Lactobacillus plantarum,L. p）、戊糖片球菌（Pediococcuspentosaceus,P. p）及其组合菌种于牦牛肉灌肠,每隔24 h测定其pH值,分别在主发酵2 d（32 ℃）,再自然风干发酵10、20、30 d后取样,采用反相高效液相色谱（reversed phase high performance liquid chromatography,RP-HPLC）对不同发酵时期牦牛肉灌肠中肽的含量进行检测和分析。结果表明,在主发酵2 d,再自然风干发酵10、20 d（除P. p与L. p＋P. p外）和30 d后,不同乳酸菌发酵剂对牦牛肉灌肠中总肽含量的影响均显著（P≤0.05）。按发酵时间由短及长的顺序,在4 个不同发酵时期检出最大总肽含量的发酵菌种依次是L. s（1.73×105 mAU·s）、P. p（1.62×105 mAU·s）、对照组（1.81×105 mAU·s）和L. p（1.83×105 mAU·s）。随着发酵时间的延长,发酵牦牛肉灌肠中的总肽含量发生了明显的变化,相对极性较弱的肽减少了,存在的主要是相对极性较大的肽;主发酵结束时,L. s发酵的处理组检出最大总肽含量,且L. s与其参与发酵L. s＋P. p和L. s＋L. p＋P. p组合中检出总肽含量的变化呈现出相似的趋势,L. s是牦牛肉灌肠中快速生成肽的良好发酵剂;在4 个发酵时期,L. p＋L. s发酵的处理组检出肽的含量均小于其单一菌种的发酵,L. p＋L. s对牦牛肉灌肠中肽的生成具有一定的拮抗作用;L. s与P. p作用不明显,但能快速降低pH值,保证产品的安全性。本实验结果为进一步生产优质牦牛肉灌肠奠定了良好的基础。     

腌腊肉制品中乳酸菌的筛选鉴定及其在腊肠中的应用

为筛选适合传统腌腊肉制品的优良乳酸菌菌株,从多种农家自制传统腌腊肉制品中分离纯化出9株优势乳酸菌。通过发酵特性筛选,得到一株性状优良菌株10M-7,并制备该菌株的干粉发酵剂,以未接种发酵剂腊肠为对照,分析此发酵剂对腊肠感官品质和微生物变化的影响。结果表明,10M-7菌株具有良好的产酸特性和抑菌性能。根据形态学、生理生化特征和16S rRNA序列分析,鉴定其为植物乳杆菌,采用冷冻干燥法制备纯种发酵剂,并制作人工发酵腊肠。发酵剂组pH值在初期便迅速下降,且始终低于对照组;发酵剂组乳酸菌迅速生长繁殖,且葡萄球菌和大肠杆菌数量与对照组相比明显降低。感官评价表明,当添加量为10~4CFU/g原料肉时,能够很好地保持和改善产品风味,使产品整体感觉更好。     

Survival of Listeria monocytogenes strains in a dry sausage model

The survival of five inoculated Listeria monocytogenes strains (DCS 31, DCS 184, AT3E, HT4E, and HR5E) was studied in dry fermented sausages prepared using two different starter cultures (starter A and B) with or without a protective Lactobacillus plantarum DDEN 2205 strain. L. monocytogenes was detected throughout ripening in every sausage sample in which the L. plantarum DDEN 2205 strain had not been used. The use of either starter A, with a high concentration of protective culture, or starter B, with a low concentration of protective culture, resulted in L. monocytogenes-negative sausages after 17 days of ripening. Differential survival was noted among the L. monocytogenes strains during fermentation. Strains AT3E and DCS 31 survived in sausages with protective cultures more often than did the other strains, whereas HT4E and HR5E were inhibited during ripening by all starter and protective cultures used. Protective cultures such as L. plantarum may be used as part of a hurdle strategy in dry sausage processing, but variations in susceptibility of different L. monocytogenes strains can create problems if other hurdles are not included.     

The occurrence and growth of microorganisms during the fermentation of fish sausage

Minced fish (mullet) sausage mixes containing added sugar, salt, nitrate, nitrite and spices were fermented (48 h, 30 degrees C) by indigenous flora or by a starter culture (Pediococcus acidilactici) and the microbial ecology and behaviour of various bacteria was monitored. Pediococcus pentosaceus and Lactobacillus plantarum dominated the indigenous fermentation, achieving populations of 10(7)-10(8) cfu/g by 48 h, and decreasing the pH of the mix to 4.5-4.7. Significant growth (10(5)-10(7) cfu/g) of Staphylococcus warneri, Staphylococcus aureus, Staphylococcus epidermidis. Micrococcus varians and Micrococcus luteus also occurred during this fermentation. Less growth was exhibited by Bacillus megaterium and yeasts. Pediococcus acidilactici dominated the fermentation when it was inoculated as a starter culture, but indigenous lactic acid bacteria (P. pentosaceus and L. plantarum) also grew to 10(7)-10(8) cfu/g. The growth of other bacteria and yeasts was restricted during fermentation with starter culture. Inoculated Escherichia coli, Salmonella typhimurium, Salmonella sofia, and Staphylococcus aureus grew to 10(6)-10(7) cfu/g in the sausage mix during indigenous fermentation. Lesser growth occurred for Bacillus cereus, Clostridium perfringens and Vibrio parahaemolyticus. Growth of these bacteria was significantly inhibited in sausage mix fermented with P. acidilactici.     

不同发酵方式对发酵马肉肠贮藏期品质劣变及食用安全性的影响

为有效确保发酵马肉肠贮藏期间的品质及安全性,研究添加汉逊德巴利酵母菌1808（Debaryomyces hansenii 1808）、木糖葡萄球菌21445（Staphylococcus xylose 21445）、植物乳杆菌E11（Lactobacillus plantarum E11）的复合发酵剂发酵组（FP）和单一乳酸菌发酵组（DZ）、自然发酵组（ZR）马肉肠贮藏过程中生物胺的变化,考察不同发酵方式对发酵马肉肠pH值、水分活度（aw）、色差、总挥发性盐基氮（total volatile basic nitrogen,TVB-N）含量、硫代巴比妥酸反应物（thiobarbituric acid reactive substances,TBARs）值及菌落总数的影响。结果表明：在贮藏期间,相比于自然发酵组和单一乳酸菌发酵组,添加复配发酵剂可有效减缓发酵马肉肠TVB-N含量、TBARs值、pH值的增幅,抑制贮藏后期有害杂菌的生长,防止产品过早发生腐败变质;此外,复配发酵剂可显著降低贮藏过程中马肉肠的水分损失（P＜0.05）,维持色泽的稳定,抑制生物胺（尸胺、腐胺、酪胺、组胺）的积累。综上,随着贮藏时间的延长,复配发酵剂可显著抑制发酵马肉肠品质的劣变及不良生物胺的过多积累。     

剁辣椒发酵过程中菌群与有机酸变化规律分析

以自然发酵为对照,以植物乳杆菌W-4进行剁辣椒纯种发酵。研究结果表明：在发酵过程中,接种发酵能使乳酸菌快速成为优势菌,较快地降低剁辣椒的pH值。接种发酵具有较高数量的菌落总数和乳酸菌,酵母菌在自然发酵和接种发酵中数量相当,变化趋势基本一致,而大肠菌群只在发酵前期出现,而后消失。有机酸测定结果表明接种发酵具有较高含量的乳酸,而自然发酵具有较高含量的柠檬酸和苹果酸,而乙酸只在自然发酵的前期出现。     

Identification of potential probiotic starter cultures for Scandinavian-type fermented sausages

Potential probiotic cultures suitable as starter cultures for the Scandinavian-type fermented sausages were identified among strains well-adapted to fermented meats as well as strains originating from a culture collection. From 15 different fermented meat products, 22 strains were isolated as dominant non-starter lactic acid bacteria (NSLAB). The isolates were identified by RAPD, API and sequence analysis of 16S rRNA and showed to be five strains of Lactobacillus sakei, five strains of Lactobacillus farciminis, five strains belonging to the group of Lactobacillus plantarum/pentosus, four strains of Lactobacillus alimentarius, two strains of Lactobacillus brevis and one strain of Lactobacillus versmoldensis. Heterofermentative strains as well as strains not growing at 37 degrees C and not lowering pH below 5.1 in a meat model were excluded leaving 9 strains for further studies. These strains together with 19 strains from a culture collection were evaluated by in vitro methods including survival upon exposure to pH 2.5 or 0.3% oxgall and adhesion to the human colon adenocarcinoma cell line Caco-2 as well as antimicrobial activity against potential pathogens. Strains that fulfilled all the probiotic criteria and showed to be fast acid producers in a meat model included three strains belonging to the group of Lb. plantarum/pentosus (MF1291, MF1298, MF1300) which originated from the dominant NSLAB of fermented meat products. MF1291 and MF 1298 were further identified as Lb. plantarum and MF1300 as Lb. pentosus. The three strains were all successfully applied as starter cultures for the production of fermented sausage. The viable count at the end of the processing period reached high cell numbers (4.7x10(7)-2.9x10(8) cfu/g) and pH of the sausages decreased to pH 4.8-4.9 without any flavour deviation compared to sausage fermented by a commercial meat starter culture.     

Effects of indigenous starter cultures on the microbial and physicochemical characteristics of Urutan, a Balinese fermented sausage

Urutan is a Balinese traditional dry fermented sausage prepared from lean pork and various kinds of spice. Urutan is different from the European sausages, because it is fermented under warm condition with fluctuating temperatures of approximately 25 degrees C at night to 50 degrees C during sun drying. In this study, two of the 71 strains of lactic acid bacteria (LAB) isolated from natural urutan fermentation were used as starter cultures: Lactobacillus plantarum U201, the dominant LAB, and Pediococcus acidilactici U318, a bacteriocin producer. A soft urutan with yellowish brown color was produced using these strains as multiple starters. The starter cultures grew in characteristic succession which reconstructed the natural fermentation process. Lactobacilli were dominant until 48 h fermentation and pediococci dominated at the later stage of fermentation. Proliferation of starter cultures produced lactic acid which resulted in the decrease in pH and coagulation of soluble protein in urutan. Both strains could eliminate the Enterobacteriaceae in urutan after 24 h fermentation, and could suppress and eliminate the occurrence of micrococci at 120 h fermentation. By using a single starter culture, no succession was observed to occur in urutan and the time of elimination of Enterobacteriaceae was delayed. Thus, the strains of L. plantarum U201 and P. acidilactici U318 have great potential for use as multiple starter cultures in urutan fermentation.     

Use of the gfp gene in monitoring bacteriocin-producing Lactobacillus plantarum N014, a potential starter culture in nham fermentation

Lactobacillus plantarum N014 is a bacteriocin-producing lactic acid bacteria originally isolated from nham, a traditional Thai fermented sausage, and in the process of development to be used as a starter culture for nham fermentation. During the fermentation process, there is a need to identify the starter culture among several naturally occurring bacteria. In this study, a new plasmid carrying the gfp (green fluorescent protein) gene was constructed based on pGKV210, an Escherichia coli/ Lactococcus shuttle vector containing an erythromycin resistance marker. The gfp gene derived from pGFPuv was placed under the control of an L-lactate dehydrogenase promoter and then inserted at the EcoRI site of pGKV210, leading to pN014-GFP. The novel plasmid was used to transform L. plantarum N014, which is a bacteriocin-producing lactic acid bacteria isolated from nham. The resulting transformant, L. plantarum N014-GFP+, was brightly fluorescent and harbored the expected plasmid. A plasmid stability test revealed that pN014-GFP was stable after 100 generations of growth under nonselective pressure. L. plantarum N014-GFP+ and its parent strain were shown to be very similar in growth rate, bacteriocin production, and lactate production. L. plantarum N014-GFP+ was able to survive in a nham model. The survival clones were still fluorescent and harbored pN014-GFP.