Dried sausages fermented with staphylococcus xylosus at different temperatures and with different ingredient levels. Part IV. Amino acid profile

Sausages with added Staphylococcus xylosus were fermented at different temperatures and with different added levels of salt, glucose, nitrite, nitrate and Pediococcus pentosaceus in accordance with a six-factor fractional design. The amounts of individual amino acids were measured and the effects of temperature and different ingredients on the amino acid profile were tested using multiple linear regression and analysis of variance. Also, the amino acid profile was correlated to the level of volatile compounds by partial least squares analysis. The study showed that the level of free amino acids was significantly affected by the different factors. High fermentation temperature and nitrite content increased the amount, while high contents of salt, nitrate, glucose and P. pentosaceus lowered the amount of free amino acids. In general, temperature and nitrate had the greatest influence. It was shown that the amounts of the volatile compounds, 2-methyl propanal, 2- and 3-methyl butanal, were inversely correlated with the amounts of valine, isoleucine and leucine, respectively, indicating that those volatiles were degradation products of the latter.     

Dried sausages fermented with Staphylococcus xylosus at different temperatures and with different ingredient levels - Part II. Volatile components

Sausages, with added Staphylococcus xylosus, were fermented at different temperatures and with different added levels of salt, glucose, nitrite, nitrate and Pediococcus pentosaceus in accordance with a six factor fractional design. The volatile compounds from the sausages were collected by dynamic headspace sampling and quantified and identified by gas chromatography and gas chromatography-mass spectrometry. The effects of temperature and different ingredients on the levels of individual volatiles were tested using multiple linear regression and analysis of variance. The study showed that sausages fermented under modern production conditions (high temperature, addition of glucose, nitrite, Pediococcus pentosaceus) in contrast to 'old-fashioned' sausages (added nitrate and fermented at low temperature) are likely to contain higher amounts of volatile acids, but lower levels of ethyl esters, certain short chain aldehydes as well as lipid autoxidation products. Several nitriles, nitro-alkanes and one organic nitrate were identified for the first time in fermented sausages. Reaction mechanisms for those compounds and other classes of compounds are proposed and discussed in detail.     

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。     

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.     

人工发酵剂对湘西腊肠微生物菌群和理化特性的影响

本文从动态角度探讨自然发酵、戊糖片球菌发酵、木糖葡萄球菌发酵和混合菌发酵腊肠在发酵过程中五种菌相(细菌总数、乳酸菌、葡萄球菌、酵母菌和肠细菌)和理化特性的阶段性变化。微生物数据显示,四组腊肠初始肠细菌数量约5.30 logCFU/g,而发酵结束,添加了戊糖片球菌腊肠组的肠细菌数量约1.00 logCFU/g,没有添加该菌腊肠组的肠细菌数量约3.30 logCFU/g,说明戊糖片球菌能有效抑制肠细菌的生长,保证了产品的安全性和稳定性;发酵初期,腊肠p H值迅速降5.30以下,到发酵中期,腊肠p H开始回升;在发酵过程中,戊糖片球菌腊肠组的TBARS(硫代巴比妥酸)值显著(p0.05)高于其他三组腊肠,而其它理化特性无显著差异。从数据分析可知,腊肠的发酵剂能有效改善其卫生质量,而对腊肠的p H值、AW和亚硝酸盐残留量没有显著影响。     

微生物发酵剂对四川风羊腿加工进程及产品特性的影响

研究微生物发酵剂对四川传统风羊腿不同加工阶段及产品特性的影响,结果表明：风羊腿的水分含量及水分活度（water activity,aw）在风干发酵初期下降较快,后期逐渐平缓;pH值在发酵开始1 周下降较快,成熟后期逐渐回升;NaCl含量逐渐增加,亚硝酸钠残留量逐渐下降;过氧化值、硫代巴比妥酸反应物值和挥发性盐基氮含量逐步上升;添加微生物发酵剂的产品水分含量、aw和pH值变化幅度更大,发酵进程更快,发酵微生物呈现出极为显著的抑制脂肪氧化和降低硝盐（硝酸钠和亚硝酸钠）残留作用,这一作用在腌制和发酵初期就已显现。     

Relationship between fat depots in carcasses of beef bulls and effect on meat colour and tenderness

The growth and the effects of four species of staphylococci and six lactic acid bacteria (LAB) of the genus Carnobacterium, Lactobacillus and Pediococcus on unsaturated free fatty acids were studied. The strains were grown in complex medium supplemented either with oleic, linoleic or linolenic acids. Growth was followed and oxidation of the substrates measured by TBARS. The strains of Staphylococcus xylosus 873, 16, Staphylococcus warneri 863 and Staphylococcus saprophyticus grew well on all the substrates. Whereas, the growth of the two strains of Staphylococcus carnosus and Staphylococcus xylosus 831 was inhibited in the media with linolenic acid. The addition of manganese to this media allowed good growth of these strains. All the LAB did not grow well in the media with linoleic acid, but their growth was favoured by addition of manganese to the media. Under our conditions, only linoleic and linolenic acids were oxidised. All the strains had no prooxidant activity. All the staphylococci limited oxidation of linoleic acid and had a small effect on linolenic acid. LAB did not limit oxidation of linoleic acid. With manganese in the media: the oxidation of the sterile controls was delayed for 2 days and then increased; strains of S. carnosus and S. xylosus inhibited oxidation of linolenic acid; and Lactobacillus plantarum and Pediococcus pentosaceus limited oxidation of linoleic acid. The two Carnobacterium, whatever the conditions, had no antioxidant properties.     

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.     

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.     

虾头自溶产物微生物混合发酵的工艺条件

以感官评定和氨基态氮含量为指标,选择最佳菌种组合。利用正交试验对发酵虾头自溶产物工艺进行优化。结果表明:最佳菌种组合为木糖葡萄球菌、戊糖片球菌和枯草杆菌。通过L9(34)正交分析试验,得出优化工艺:发酵温度30℃,接种量为3%,发酵时间8 h。在该工艺条件下,发酵液虾味明显,发酵味增强,氨基态氮含量达到0.453 mg/100 g。     

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.     

The pH-unrelated influence of salt, temperature and manganese on aroma formation by Staphylococcus xylosus and Staphylococcus carnosus in a fermented meat model system

The influence of manganese (0.01-0.1-1.0 microg/g), temperature (15-24 degrees C) and salt (3-4% w/w) on volatile formation in model minces inoculated with Pediococcus pentosaceus and either Staphylococcus xylosus or Staphylococcus carnosus was studied in a full factorial experiment. In order to study the direct, pH-unrelated effect of the parameters, data were analysed by use of multiple linear regression and partial least-squares regression both before and after transformation of the volatile responses into pH-orthogonal (pH-unrelated) responses. By using the pH-orthogonalised data, the overall interpretability of the experiment was increased, and new cause-and-effect relations were suggested. Approximately 50% of the total variance in volatile levels was due to differences caused by S. xylosus and S. carnosus, and another 30% was related to differences in pH development. The remaining 20% covered pH-orthogonal effects of manganese, temperature and salt plus the experimental noise. From this, it was concluded that most of the variation in volatile profiles caused by manganese, temperature and salt was in fact directly or indirectly caused by changes in lactic acid bacterial activity and pH.     

一株戊糖片球菌在低盐固态酱油工艺中的应用

酱油发酵酱醅中筛选到一株戊糖片球菌,对该菌的生长温度、生长pH值、食盐耐受度等生长特性进行研究。采用离子交换色谱法对该菌株产有机酸的情况进行测定;将该菌株添加到低盐固态酱油后酵过程中,以不添加乳酸菌的实验组作对照,对酱油样品理化指标、经济指标进行分析,并采用顶空固相微萃取-气相色谱-质谱法(HS-SPME-GC-MS)对酱油挥发性香气成分进行对比分析。结果表明,该乳酸菌的生长条件与酱油酱醅发酵环境相适应,其发酵产酸以乳酸和乙酸为主,与酱油中有机酸主成分相一致,将该菌株用于低盐固态酱油发酵,对酱油香气和其他指标均有不同程度的促进作用。     

发酵剂对发酵香肠食用品质的影响及其在不同直径香肠中的应用

以商业发酵剂（木糖葡萄球菌＋戊糖片球菌）为对照组,不同发酵剂组合（木糖葡萄球菌＋副干酪乳杆菌、木糖葡萄球菌＋戊糖片球菌＋副干酪乳杆菌）为实验组,通过对发酵香肠水分含量、pH值、水分活度、色泽、质地、风味和感官品质等指标进行测定,确定最佳发酵剂及适宜发酵的香肠直径。结果表明：相较于商业发酵剂,木糖葡萄球菌与副干酪乳杆菌组合发酵的香肠总体可接受性相对较高,且庚醛、1-辛烯-3-醇、乳酸乙酯、戊酸乙酯、癸酸乙酯、2-甲基丙酸乙酯等愉悦风味物质为该组独有,其特征主要表现为更加浓郁的清新味、甜香味、果香味和花香味;适宜的直径（21 mm）、水分含量（（25.40±0.00）%）和硬度（（2 812.46±767.93）g）以及相对较高的pH值（pH 5.57±0.02）是该组发酵香肠口感显著高于其他2 组的重要原因。因此,木糖葡萄球菌与副干酪乳杆菌组合发酵的小直径香肠食用品质最佳。     

威宁火腿中微生物的分离鉴定及其耐受特性

以贵州威宁火腿为研究对象,对其中的微生物开展分离鉴定及耐盐、耐亚硝酸盐和耐低温特性研究。结果表明：共分离得到10 株菌,包括4 株马胃葡萄球菌（Staphylococcus equorum）、2 株木糖葡萄球菌（Staphylococcus xylosus）、2 株乳酸片球菌（Pediococcus acidilactici）、1 株变平滑假丝酵母菌（Candida metapsilosis）和1株近平滑假丝酵母菌（Candida parapsilosis）;其中,马胃葡萄球菌WN1、变平滑假丝酵母菌WN9和近平滑假丝酵母菌WN10耐受9% NaCl、150 mg/kg亚硝酸盐和4 ℃低温条件的能力较强。     

发酵香肠中生物胺生成量的菌株效应分析

本实验对乳杆菌(米酒乳杆菌和戊糖乳杆菌)和片球菌(戊糖片球菌和小片球菌)在发酵香肠中生物胺的生成量进行了分析。结果表明,供试乳杆菌和片球菌均能产生不等量的组胺、尸胺、腐胺、酪胺、亚精胺和精胺,但都不产生色胺,产品中的生物胺生成量表现出明显的菌株效应。采用复配菌种(片球菌+葡萄球菌)发酵可以明显降低香肠中组胺、尸胺、腐胺和酪胺以及总生物胺的含量。     

Evaluation of different starter cultures (Staphylococci plus Lactic Acid Bacteria) in semi-ripened Salami stuffed in swine gut

The ripening properties were evaluated in semi-ripened pork Salami started by different cultures: CXP (Pediococcus pentosaceus, Staphylococcus xylosus and Staphylococcus carnosus); RAP (Lactobacillus plantarum, Lactobacillus sakei, S. xylosus and S. carnosus); and GY2 (L. sakei, S. xylosus and S. carnosus). These starters were chosen by their technological activities: CXP (rapid acidifying); RAP (medium acidifying); and GY2 (slow acidifying plus intense reddening-flavouring). Salami was enhanced with Ponceau 4R red and sodium glutamate, stuffed in pig gut and aged for 12 days at 15 °C/65-85%RH. Dehydration, reddening, proteolysis, fat acidity and lipid oxidation were hardly affected by the starters. Acidification by LAB strongly influenced the flavour. L. sakei plus L. plantarum provided a better flavouring than a low dose of L. sakei, and, especially, than over-acidifying P. pentosaceus. A final pH of 5 seems to be suitable for preventing aroma and taste defects in semi-ripened Salami. Spontaneous lactic microflora showed great acidifying potential.     

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。     

食盐对传统发酵肉成熟过程中微生物菌群、理化性质及盐溶性蛋白特性的影响

以不同食盐添加量（0%、1%、3%、5%、7%）发酵肉（戊糖片球菌∶木糖葡萄球菌＝1∶1）为研究对象,测定发酵肉微生物菌群、pH值、水分含量、亚硝酸盐残留量,以及最终成熟发酵肉中食盐、总氮、非蛋白氮及盐溶性蛋白含量,研究食盐添加量对发酵肉成熟过程中微生物菌群、理化性质及盐溶性蛋白特性的影响。结果表明：随着食盐添加量的增加,微生物菌群数量基本呈先上升后下降的趋势;食盐添加量3%的发酵肉,蛋白质水解指数最高,盐溶性蛋白含量最低,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳条带最为清晰。