鲊肉粉中乳酸菌和葡萄球菌的筛选及鉴定

从传统发酵鲊肉粉中分离、筛选发酵性能优良的乳酸菌和葡萄球菌,为鲊肉粉接种发酵提供理论依据。按照肉制品发酵菌株的筛选标准,利用形态学特征和16S rDNA序列分析鉴定菌株,筛选出2株乳酸菌A1、C7和2株葡萄球菌S6、S7。结果表明:乳酸菌菌株A1、C7均为植物乳杆菌(Lactobacillus plantarum),能快速产酸,具有较好抑菌特性;葡萄球菌菌株S6为沃氏葡萄球菌(Staphylococcus warneri),S7为巴氏葡萄球菌(Staphylococcus pasteuri),能产蛋白酶和脂肪酶,具有良好的耐盐性和耐亚硝酸盐性,对不同抗生素有不同的敏感性。筛选得到的4株菌株具有良好的发酵特性,乳酸菌和葡萄球菌之间无拮抗作用,复配后用可于鲊肉粉接种发酵。     

风味咸鱼中乳酸菌和葡萄球菌的分离与鉴定

为优化咸鱼的加工工艺,对优质北海淡口咸鱼进行乳酸菌和葡萄球菌的分离和鉴定。分别采用MRS和MSA培养基,从咸鱼中分离出乳酸菌13 株和葡萄球菌10 株。经生理生化实验初筛出乳酸菌3 株、葡萄球菌两株。初步鉴定乳酸菌为戊糖片球菌,两株葡萄球菌分别为肉糖葡萄球菌和木糖葡萄球菌。     

应用多重PCR快速鉴定自然发酵肉制品中6 种葡萄球菌

肉葡萄球菌（Staphylococcus carnosus）、木糖葡萄球菌（S. xylosus）、腐生葡萄球菌（S. saprophyticus）、表皮葡萄球菌（S. epidermidis）、马胃葡萄球菌（S. equorum）和松鼠葡萄球菌（S. sciuri）是发酵肉制品中常见的葡萄球菌,为准确、快速地检测和鉴定这些菌种,建立6?种葡萄球菌的多重聚合酶链式反应（polymerase chain reaction,PCR）方法并进行验证。以上述菌种的gap、rpoB、SodA和SNc为靶基因,设计和筛选菌种特异性引物6?对,优化PCR体系,建立6?种葡萄球菌的多重PCR鉴定方法。实验结果显示多重PCR的基因组DNA检测灵敏度可达到4?pg,活菌检测灵敏度可达到2×102?CFU;采用建立的多重PCR方法对传统肉制品中分离的8?株葡萄球菌进行鉴定,结果与16S rDNA序列分析、生理生化鉴定结果一致。建立的多重PCR方法具有较高的种间特异性,可快速、准确地用于发酵肉制品中肉葡萄球菌、木糖葡萄球菌、表皮葡萄球菌、马胃葡萄球菌、松鼠葡萄球菌和腐生葡萄球菌的检测和鉴定,具有较好的应用前景。     

从自然发酵风干肠中分离出的葡萄球菌生产性能的研究

从自然发酵风干肠中分离出五株葡萄球菌,通过菌种的耐亚硝酸盐能力、耐酸能力、生长温度和最适生长温度、蛋白质及脂肪降解活性等实验,对其生产性能进行研究.结果表明,木糖葡萄球菌(Staphylococcus xylosus,S2)能在0.015‰亚硝酸盐中生长;耐pH为3的酸性环境;不耐65℃的高温,烘烤后可被灭活;30～35℃内旺盛生长;具有降解蛋白质和脂肪的能力,符合作为肉制品发酵剂的要求.     

发酵肉用耐高温产香葡萄球菌优质菌株的筛选

为了开发发酵肉制品高温加工新工艺,首先需要获得新型耐高温肉用发酵剂。因此,从湘西腊肉中取样,通过耐盐性、耐酸性、产酶活性、硝酸盐还原能力等指标筛选出4株耐高温葡萄球菌S5、S9、S10和S20,鉴定为1株表皮葡萄球菌,2株华纳氏葡萄球菌,1株木糖葡萄球菌。该些菌株的耐盐、耐亚硝酸盐、耐高温和产香能力较强,有望在发酵肉制品生产中使用。     

肉品发酵剂葡萄球菌和微球菌的分离筛选及其与乳酸菌混合培养的研究

本试验采用MSA培养基从二十多种传统发酵肉制品中分离菌株198株，通过对分离菌株发酵特性试验，筛选出两株过氧化氢酶阳性球菌，经过初步鉴定为肉糖葡萄球菌(Staphylococcus carnosus)S15和S113，并研究了戊糖片球菌(Pediococcus pentosaceus)P20、米酒乳杆菌(Lactobacillus sake)J17分别同两株葡萄球菌在模拟肉汤中混合培养的相互关系，初步确定了肉品混合发酵剂P20 S15。     

不同发酵香肠中微球菌与葡萄球菌的筛选与鉴定

144株分离色拉米发酵香肠的微球菌科菌株被鉴定.所用的发酵香肠来自三个地区:新疆、山东、湖南,每个地区选择两个厂,香肠出自制作的的三个不同阶段.其中木糖葡萄球菌是主要菌种(95%),经过生化反应鉴定出共有12种典型的木糖葡萄球菌,2号和5号两种菌占大多数(38.8%,33.6%).根据观察,木糖葡萄球菌5号,显示硝酸盐还原酶和脲酶活性,适度的蛋白质和脂肪分解能力,产生乙偶姻,适合做发酵剂.     

肉制品发酵剂木糖葡萄球菌J23发酵条件的研究

高质量的发酵肉专用发酵剂是工业化生产发酵肉的关键。本研究从金华火腿中分离鉴定了可作为发酵剂的木糖葡萄球菌J23,并对其发酵条件(包括5L罐放大试验)、冷冻干燥条件、浓缩培养、菌株安全性等方面进行了研究。结果说明:在最适培养温度30℃下,2#简单培养基为最佳培养基,用2 mol/l的NaOH作为中和剂控制发酵pH7.3,发酵周期为20h,木糖葡萄球菌J23活菌数可达到3.4×109CFU/mL。冷干保护剂以蔗糖效果最好,冷干后活菌数能达到4×1010CFU/g。发酵剂菌株经动物毒性试验表明属无毒级微生物。     

发酵香肠葡萄球菌的生理生化特性的测定

试验采用中国传统发酵香肠作为采集样品,从中分离纯化出具有革兰氏阳性和触酶阳性的葡萄球菌,对此菌株做生理生化特性鉴定,以对发酵香肠的菌株进行进一步的筛选,从中选出不产粘液、发酵葡萄糖不产气、不产色素、不产H2S、不具有氨基酸脱羧酶、不产氨、15℃能还原硝酸盐、pH4.5条件下生长的菌株作为发酵香肠中的优良葡萄菌株,最后测定优势菌株24小时的生长曲线和pH变化曲线。     

木糖葡萄球菌添加量对发酵牛肉串品质特性的影响

研究木糖葡萄球菌的添加量对发酵牛肉串理化性质和感官特性的影响。将不同添加量的木糖葡萄球菌分别与干酪乳杆菌(107CFU/g)复配作为发酵剂用于生产发酵牛肉串,测定发酵牛肉串在发酵成熟过程中的pH值、水分活度、颜色、质构及感官指标的变化。结果表明：添加108CFU/g木糖葡萄球菌的发酵肉串气味值最高(4.75±0.38),但木糖葡萄球菌菌种添加量为106、108CFU/g时,发酵肉串的色泽、质地、总体可接受性均低于木糖葡萄球菌菌种添加量为107CFU/g的肉串;而原料肉中添加107CFU/g木糖葡萄球菌的实验组在发酵终点时产品获得优良的感官特性(总体可接受性达到最高值5.31),pH值达到5.4,质构特性好,并与其他实验组存在显著性差异(P≤0.05)。     

Generation of flavour compounds in fermented sausages-the influence of curing ingredients, Staphylococcus starter culture and ripening time

The volatile profiles of fermented sausages made with either Staphylococcus xylosus or Staphylococcus carnosus starter cultures were studied with regard to the influence of salt concentration, ripening time and three different combinations of curing ingredients-nitrate, nitrite or nitrite/ascorbate. Emphasis was laid on volatile compounds originating from degradation of branched-chain amino acids. Volatile compounds were collected using dynamic headspace sampling and were identified by gas chromatography/mass spectrometry (GC/MS). Development in water activity, water loss and pH was monitored throughout maturation. Curing salts had a pronounced effect on the level of volatile compounds. In particular, curing with nitrate instead of nitrite resulted in a striking difference. Generally, nitrate increased the level of volatile compounds compared to nitrite, whereas ascorbate had only a small influence. The concentration level of NaCl had a considerable effect on the amount of volatile compounds but the effect was highly related to the ripening stage. Most compounds, but not all, increased in concentration as ripening proceeded. Major differences in the development of volatile compounds were observed depending on whether S. xylosus or S. carnosus were used as starter culture. In particular the effects of nitrate was much more predominant in the sausages made with S. carnosus than S.xylosus.     

不同乳酸菌发酵剂对发酵红肠品质的影响

为研究乳酸菌发酵对红肠品质的影响,将发酵技术应用于本无发酵工艺的红肠制品中,筛选出能够提高红肠品质的乳酸菌发酵剂。分别将常应用于发酵肉制品的7 种商业乳酸菌发酵剂（木糖葡萄球菌＋戊糖片球菌（THM-17）、木糖葡萄球菌＋清酒乳杆菌＋类植物乳杆菌（PRO-MIX5）、木糖葡萄球菌＋肉葡萄球菌＋清酒乳杆菌（WBL-45）、木糖葡萄球菌＋戊糖片球菌＋植物乳杆菌（VHI-41）、木糖葡萄球菌＋戊糖片球菌＋植物乳杆菌（SHI-59）、肉葡萄球菌＋木糖葡萄球菌（WBX-43）和戊糖片球菌＋木糖葡萄球菌＋肉葡萄球菌＋乳酸片球菌（VBM-60））及8 种单菌（弯曲乳杆菌、戊糖乳杆菌、清酒乳杆菌-1、戊糖片球菌、木糖葡萄球菌、肉葡萄球菌、清酒乳杆菌-2、植物乳杆菌）以107 CFU/g的接种量接种至腌制后的肉馅中,拌馅灌肠后于35 ℃、80%湿度条件下发酵12 h,取样测定发酵后样品的乳酸菌数和细菌总数,再经干燥、蒸煮、烟熏、烘烤制得成品,测定其感官、pH值、色差、质构、亚硝酸盐、硝酸盐、生物胺及N-亚硝胺含量等指标。结果表明：15 种发酵剂中以木糖葡萄球菌和植物乳杆菌2 种乳酸菌发酵剂应用效果较好,所制得产品pH值分别为5.26和5.04,色泽美观,弹性适中,亚硝酸盐残留量（10.84、10.13 mg/kg）低,可显著抑制N-亚硝胺的形成（N-二甲基亚硝胺含量分别为1.29、2.51 μg/kg）,生物胺总量较低。由此说明,木糖葡萄球菌和植物乳杆菌能够显著提高红肠产品的安全品质。     

Characterization and technological properties of Staphylococcus xylosus strains isolated from a Tunisian traditional salted meat

The technological properties of strains of Staphylococcus xylosus were studied to select the most suitable for use as starter cultures for the production of dried fermented meat products. Strains of S. xylosus were isolated from traditional salted Tunisian meat and were identified by biochemical and molecular methods. Thirty strains of S. xylosus were studied to evaluate their catalase, nitrate reductase, lipolytic, proteolytic and antibacterial activities as well as growth ability at different temperatures, pH's and NaCl concentrations. All strains of S. xylosus had catalase activity and were able to reduce nitrates to nitrites. The nitrate reductase activity increased when the strains were kept under anaerobic conditions. Proteolytic activity on milk and on gelatin agar was demonstrated for 100% and 83.3% of the S. xylosus isolates, respectively. However extracellular proteolytic activity as assessed by the azocasein method was poor in all the strains. Lipolytic activity as assessed by the agar method showed that 76.6% of strains of S. xylosus could hydrolyze Tween 20 against 33.3% that could hydrolyze tributyrin. Tween 80 was hydrolyzed by only 10% of strains. Strains of S. xylosus hydrolyzed pork fat better than beef and lamb fat. The majority of strains had antibacterial activity against Salmonella arizonae, Staphylococcus aureus, Pseudomonas aeuroginosa, Escherichia coli and Enterococcus faecalis.     

Growth and aroma production by Staphylococcus xylosus, S. carnosus and S. equorum--a comparative study in model systems

A laboratory medium inoculated with 20 different Staphylococcus strains was prepared in accordance with a full factorial experimental design investigating the effect of temperature, pH, NaCl and glucose on growth. The 12 strains most suited to growth in a fermented meat environment were inoculated in sausage minces together with Pediococcus pentosaceus, incubated at 25 degrees C for 1 week and the produced aroma compounds collected. The data were analysed by multiple linear regression and partial least squares regression analysis. The results showed that increasing pH and temperature from 4.6 to 6.0 and 10 to 26 degrees C, respectively, increased growth of all strains with strong synergy between temperature and pH. Increasing salt concentration from 5% to 15% w/v decreased growth of most strains, but the effect of pH and temperature was much stronger than the effect of salt. Strains of S. carnosus were more salt tolerant than strains of S. equorum and S. xylosus, especially at high pH and temperature. Addition of glucose up to 0.5% w/v had no significant influence on growth of any of the strains. With regard to aroma production, species characteristics were detected. S. carnosus and S. xylosus were quite different regarding the overall aroma profiles, whereas the profiles of S. equorum lied somewhere in-between. Contrary to S. carnosus, S. xylosus and S. equorum did not produce 2-methyl-1-butanol. On the other hand, in particular, S. xylosus produced more 3-methyl-1-butanol. Except for one of the strains of S. equorum, S. xylosus and S. equorum formed more diacetyl, 2-butanone and acetoin and also more of the methyl-branched ketones arising from degradation of leucine, isoleucine and valine. S. carnosus produced more methyl-branched aldehydes, acids and corresponding esters from leucine, isoleucine and valine compounds that have been correlated with fermented sausage maturity in former studies. S. equorum produced the least of the methyl-branched aldehydes.     

Effect of ascorbate, nitrate and nitrite on the amount of flavour compounds produced from leucine by Staphylococcus xylosus and Staphylococcus carnosus

Resting cells of Staphylococcus xylosus and S. carnosus were incubated with ascorbate, nitrate and nitrite in defined reaction medium and their degradation of (3)H-labelled leucine into methyl-branched catabolites were studied using HPLC/radiometric detection. The experiments were carried out with and without addition of α-ketoglutarate. The main catabolic product of leucine degradation was 3-methylbutanoic acid but also small amounts of α-hydroxy isocaproic acid were produced. Nitrite addition lowered the concentration of 3-methylbutanoic acid for both Staphylococcus species and this effect was strongly amplified by ascorbate for S. xylosus but not for S. carnosus. For both species ascorbate alone had little if any effect. Also nitrate lowered the concentration of 3-methylbutanoic acid for S. xylosus. The concentration of α-hydroxy isocaproic acid was, however, increased by addition of nitrite and nitrate for S. xylosus. Addition of α-ketoglutarate generally increased the concentration of 3-methylbutanoic acid for both S. xylosus and S. carnosus.     

PROTEOLYTIC ACTIVITY OF BACTERIAL STARTER CULTURES FOR MEAT FERMENTATION

Proteolytic activity of fermented meat starter cultures Pediococcus acidiliactic, Lactobacillus sake, Lactobacillus curvatus, Streptomyces griseus, Staphylococcus xylosus and Staphylococcus carnosus was examined after inoculation of sterile beef protein extracts containing 1% dextrose. Analysis for primary amines in extracts generated during fermentation of glucose showed that L. sake had the highest (P<0.05) proteolytic activity. The other cultures ranked from highest to lowest activity as follows : L. curvatus < S. carnosus < S. griseus < P. acidilactici < S. xylosus. All inoculated extracts showed a pH decline by 96 h to pH 3.76-4.31 whereas the control extract remained at 6.0-5.95. All cultures except S. xylosus had CFU increases within 24 h of inoculation. Only L. sake and S. xylosus had counts less than their initial level after 96 h. The results provide evidence that these commercially available starters for meat fermentation do possess significant proteolytic activity when tested using beef protein extracts.     

芹菜粉对天然中式香肠颜色影响及工艺优化的研究

天然中式发酵香肠是不添加人造着色成分、化学防腐剂或任何其他人造或合成成分的一种产品。本文以芹菜粉、产硝酸还原酶的木糖葡萄球菌和肉糖葡萄球菌作为天然配料,以中式香肠的红度值a和亚硝酸盐残留量为指标,通过单因素实验确定了四因素(接种量、菌种配比、发酵温度和发酵时间)的优工艺参数的基础上,采用正交实验L9(34)的方法,确定最优的发酵工艺参数条件为:A1B3C2D2,即发酵时间为30h,发酵温度为30℃,发酵菌株接种量为107cfu/g,发酵菌种配比采用木糖葡萄球菌:肉糖葡萄球菌=1:2。     

木糖葡萄球菌和肉葡萄球菌混合发酵剂对腊肉品质的影响

以不添加发酵剂的腊肉为空白对照,从理化特性、微生物分析、感官评价、香气活性化合物组成及电子鼻分析角度,研究木糖葡萄球菌和肉葡萄球菌混合发酵剂对腊肉品质的影响。结果表明,添加葡萄球菌混合发酵剂对产品的品质影响较大。与空白组比,葡萄球菌混合发酵剂可显著提高产品的酸价（2.12→3.28?mg/g）、蛋白质降解指数（16.32%→19.24%）、红度值a*（9.25→11.08）、香气和接受度得分及氨基酸代谢源香气活性化合物的种类和含量（3-甲基丁醛、3-甲基丁酸）,同时显著降低产品的过氧化值（0.15→0.07?g/100?g）、硫代巴比妥酸反应物含量（0.25→0.17?mg/kg）和脂肪氧化源香气活性化合物的含量（己醛和壬醛）,而对成品的基本理化指标（水分含量、水分活度、pH值）没有显著影响。电子鼻传感器能够将2?组产品区分开,说明加入发酵剂后,腊肉的风味有所改变。因此,通过添加木糖葡萄球菌和肉葡萄球菌混合发酵剂可以显著促进腊肉中蛋白质和脂肪的水解、改善产品的色泽,促进发酵风味快速形成,同时延缓脂肪氧化。