Impact of Lactic Acid Bacteria Screening on the Quality of Fermented Sausage
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摘要: 利用香肠模型结合主成分分析(PCA),筛出了一株对香肠风味物质有显著提升的植物乳杆菌M-25(Lactobacillus plantarum,M-25)。将其添加到香肠中,以未接菌组作为对照,在川式香肠制作的第0、1、3、7、14 d对pH、Aw、色泽、过氧化值(POV)、硫代巴比妥酸(TBARS)以及挥发性风味物质进行了检测。应用偏最小二乘模型(PLS-DA),以VIP(投影中的变量重要性)>1,P<0.05作为筛选条件对香肠中的特征风味物质进行评价。结果表明,随发酵时间推移对照组和接种植物乳杆菌香肠的pH均先降低后缓慢上升;Aw显著降低(P<0.05);POV和TBARS显著升高(P<0.05);而色泽却无显著性差异(P>0.05)。与对照组相比,接种植物乳杆菌香肠的pH、Aw、POV和TBARS值均明显更低,且接种植物乳杆菌香肠中醇类、酮类和酯类物质均明显高于对照组,能明显提升特征风味物质(异戊酸、3-羟基-2-丁酮、苯乙醛和苯乙醇)的含量。综上所述,利用香肠模型筛选的植物乳杆菌M-25能有效提升香肠品质和风味特性。Abstract: Utilizing a sausage model and principal component analysis (PCA), a strain of Lactobacillus plantarum that significantly improved the flavor substance of sausage was selected. It was added to the sausage, and the pH, Aw, color, peroxide value (POV), thiobarbituric acid (TBARS), and volatile flavor substances of Sichuan sausage production were measured at 0, 1, 3, 7 and 14 d, using the uninoculated group as control. A partial least squares discriminant analysis model (PLS-DA) was applied to evaluate the characteristic flavor substances in sausages with VIP (variable important in projection) > 1 and P < 0.05 as screening conditions. Results showed that, the pH of control and Lactobacillus plantarum-fermented sausages declined and then gradually rose with fermentation time. The Aw reduced dramatically, the POV and TBARS increased significantly, while there was no significant change in color (P>0.05). And the pH, Aw, POV and TBARS values were significantly lower in Lactobacillus plantarum inoculated sausages compared to the control group. In comparison to the control group, the sausages inoculated with Lactobacillus plantarum had significantly higher concentrations of alcohols, ketones, and esters, as well as significantly enhanced the concentrations of characteristic flavor substances (isovaleric acid, 3-hydroxy-2-butanone, phenylethylaldehyde and phenylethanol). In summary, Lactobacillus plantarum M-25 selected by the sausage model was effective in enhancing the quality and flavor characteristics of sausages.
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表 1 16 d模型中风味化合物含量
Table 1. Flavor compounds in the 16 d model
序号 名称 绝对含量(μg/kg) CT M-2 M-3 M-11 M-25 1 异戊醇 ND 22.06±1.06 95.03±9.36 221.48±10.32 ND 2 2,3-丁二醇 413.84±26.04 401.72±30.26 1036.12±98.26 408.94±10.65 530.79±23.65 3 2-乙基己醇 10.78±1.65 9.84±1.32 ND ND ND 4 2-丁基-1-辛醇 ND 11.59±1.22 ND ND ND 5 苯乙醇 ND ND ND 38.46±4.68 ND 6 3-甲基-1-丁醇 111.76±16.76 ND ND ND 194.32±13.64 7 辛醇 7.74±1.33 ND ND ND ND 醇类 544.12±45.78 445.21±33.86 1131.15±107.62 668.88±20.97 725.11±37.29 8 2-戊酮 ND 23.24±2.36 ND 97.38±12.32 52.03±10.98 9 2-庚酮 ND 62.97±2.68 19.78±3.68 232.34±44.36 117.58±16.68 10 3-羟基-2-丁酮 1286.56±100.36 1036.45±131.32 2071.75±266.22 2644.92±246.35 2626.72±232.38 11 5-甲氧基-2-戊酮 ND 14.50±1.36 ND ND ND 12 2-壬酮 ND 59.35±8.92 ND 255.99±43.23 184.07±10.35 13 异佛尔酮 28.37±8.32 8.63±2.35 33.36±2.36 ND 51.43±4.65 14 5-甲基-2-己酮 12.29±1.33 ND 18.28±2.26 ND ND 15 甲基壬基甲酮 ND ND ND 121.61±10.23 172.62±16.75 16 邻甲基苯乙酮 ND ND ND 61.04±6.12 ND 17 3-甲基-2-丁酮 ND ND ND ND 49.49±3.26 18 2-十三烷酮 ND ND ND ND 27.77±2.28 酮类 1327.22±110.01 1205.14±148.99 2143.17±274.52 3413.28±362.61 3281.71±297.33 19 丁酸 ND 12.89±0.99 ND ND 32.64±6.73 20 2-甲基丁酸 ND ND ND 18.17±1.33 71.90±18.36 21 异戊酸 ND ND ND ND 21.27±3.65 酸类 ND 12.89±0.99 ND 18.17±1.33 125.81±28.74 22 癸酸乙酯 25.97±3.33 ND 35.05±4.33 58.62±4.85 63.99±16.32 23 辛酸乙酯 ND ND ND ND 30.86±6.33 酯类 25.97±3.33 ND 35.05±4.33 58.62±4.85 94.85±22.65 24 二甲基三硫 ND 12.40±1.05 ND 54.80±4.66 174.52±10.32 25 二甲基二硫 ND ND 863.41±132.25 3477.62±289.35 3643.63±352.32 硫化物 ND 12.40±1.05 863.41±132.25 3532.42±294.01 3818.15±362.64 26 苯甲醚 ND ND ND 224.32±43.38 109.29±32.33 27 对甲苯甲醚 ND ND ND 27.52±2.35 ND 醚类 ND ND ND 251.84±45.73 109.29±32.33 28 1,2,4,5-四甲苯 25.79±3.62 14.47±1.33 ND ND ND 29 5-异丙基间二甲苯 8.04±1.02 5.61±1.21 ND ND ND 30 十二烷 40.17±6.87 17.82±4.33 28.73±4.05 ND 36.62±4.36 31 2,4-二甲基癸烷 24.24±1.22 ND 14.27±1.33 ND ND 32 叔丁基-苯 ND ND 19.76±1.62 ND ND 33 邻异丙基甲苯 ND ND ND ND 98.60±23.37 34 2,6-二甲基十一烷 7.48±1.03 ND ND ND ND 35 4,8-二甲基十一烷 6.65±0.98 ND ND ND ND 烷烃类 112.37±14.74 37.9±6.87 62.76±7 ND 135.22±27.73 36 吲哚 ND ND 12.34±1.32 ND ND 37 苯酚 ND ND ND ND 363.74±44.35 其他 ND ND 12.34±1.32 ND 363.74±44.35 主要风味成分(醇类、酮类、酸类、酯类和硫化物) 1897.31±155.79 1675.64±185.94 4172.78±518.72 7691.37±683.77 8045.63±748.65 共计 2009.68±189.63 1713.19±168.06 4259.51±264.35 7943.21±436.26 8752.48±497.63 注:“ND”表示未检测。 表 2 M-25菌株16S rDNA相似性分析
Table 2. 16S rDNA similarity analysis of M-25 strain
菌株 对比菌株 登录号 同源率(%) M-25 Lactiplantibacillus plantarum JCM8341 LC633333.1 100 M-25 Lactiplantibacillus plantarum MSD1 CP050257.1 100 M-25 Lactiplantibacillus plantarum DW12 CP058967.1 100 M-25 Lactiplantibacillus plantarum CHE37 MZ571407.1 100 M-25 Lactiplantibacillus plantarum GR1187 CP078132.1 100 M-25 Lactiplantibacillus plantarum GR1184 CP078129.1 100 M-25 Lactiplantibacillus plantarum XJ25 CP068448.1 100 M-25 Lactiplantibacillus plantarum GVS1 MZ404900.1 100 M-25 Lactiplantibacillus plantarum ATCC202195 CP063750.1 100 表 3 香肠色度的变化
Table 3. Change of sausage coloration
项目 分组 时间(d) 0 1 3 7 14 L* CL 62.72±3.09Bb 59.29±8.96Aab 52.56±6.66Aa 47.39±0.95Aa 43.18±8.2Aab LP 64.48±7.23Aa 58.31±5.4Ab 53.44±8.43Aab 46.83±3.4 Aab 40.80±4.67Aa a* CL 23.08±3.13Aa 17.15±4.15 Aab 11.04±0.9Abc 9.38±1.96Ac 7.02±4.8Ac LP 19.63±3.19 Aa 19.50±4.25 Aa 11.70±1.94 Ab 6.99±4.08 Ab 7.15±1.95Ab b* CL 23.08±3.13Aa 11.88±4.37 Ab 9.54±1.6 Ab 7.44±1.6 Ab 11.24±1.91 Ab LP 18.24±1.58Aa 15.93±6.32Aa 5.75±1.97 Ab 8.48±1.64 Ab 8.6±1.68 Ab 注:A-B不同大写字母表示L*、a*、b*分别在相同时间的不同处理香肠差异显著(P<0.05);a-d不同小写字母表示相同处理组在不同时间的差异显著(P<0.05)。 表 4 香肠中风味物质绝对含量(μg/kg)
Table 4. Absolute content of flavor substances in sausages (μg/kg)
序号 化合物名称 CL LP 0 d 1 d 3 d 7 d 14 d 0 d 1 d 3 d 7 d 14 d 1 己醇 501.45±20.36 ND ND 111.81±9.77 ND ND ND ND ND ND 2 庚醇 7.49±2.60 20.09±5.33 ND 6.10 ND 20.22±3.78 16.15±7.67 ND 7.69±1.87 ND 3 (2R,3R)-(-)-2,3-丁二醇 ND 165.89±0.97 599.95±54.89 ND ND ND 116.22±14.35 186.18±31.22 ND 37.24±3.50 4 1-辛烯-3-醇 98.92±11.34 417.66±48.3 190.52±11.58 275.84±3.05 274.06±22.39 414.67±24.75 374.55±47.09 128.18±23.75 276.68±41.67 327.81±29.04 5 4-乙基环己醇 ND 18.07±7.31 ND ND ND 13.82±2.78 18.94±6.78 ND ND ND 6 反式-2-辛烯-1-醇 ND ND ND ND ND 8.98±1.35 16.36±3.88 ND ND ND 7 4-甲基-5-癸醇 21.84±4.88 126.86±23.01 17.13±2.43 ND ND 22.36±1.81 87.68±9.13 ND ND 45.62±4.51 8 苯乙醇 ND 37.97±17.54 41.31±5.14 ND ND ND ND 45.35±5.11 81.96±3.43 252.34±6.27 9 戊醛 8.63±3.44 3.13±1.42 ND 43.89±1.70 40.81±1.79 6.67±1.11 2.30±1.06 ND 40.02±8.71 ND 10 己醛 30.61±8.67 265.22±36.4 ND 1737.45±164.2 1210.23±61.85 167.17±14.75 34.77±9.18 18.32±5.90 1543.60±69.15 401.02±47.31 11 庚醛 ND ND ND 79.45±0.98 83.45±2.13 25.45±2.27 18.03±1.38 12.90±1.24 103.41±23.04 54.65±1.81 12 苯乙醛 ND 97.42±14.73 39.11±1.04 ND ND 923.80±27.10 199.31±34.45 42.29±7.34 ND 34.96±9.44 13 3-甲硫基丙醛 ND ND ND ND ND 15.85±1.63 ND ND ND ND 14 反-2-辛烯醛 ND 22.95±2.84 ND 40.19±2.44 41.92±3.53 41.30±3.63 29.05±1.94 ND 52.59±9.69 ND 15 壬醛 65.72±14.43 64.73±16.1 9.75±3.80 102.60±0.21 102.57±2.54 104.55±6.65 ND 25.60±4.70 167.12±8.19 78.34±3.07 16 (E,E)-2,4-壬二烯醛 17.42±4.63 ND 11.98±3.95 49.87±3.12 18.86±3.32 ND ND ND ND 26.94±8.10 17 反式-2-壬烯醛 ND ND ND ND ND 6.34±1.49 2.43±0.43 ND ND ND 18 桃醛 ND ND ND ND ND 87.82±1.35 ND ND ND ND 19 2-庚酮 ND 38.91±6.95 40.37±6.80 ND ND 12.52±1.85 21.07±1.80 27.85±6.66 ND ND 20 3-羟基-2-丁酮 16.20±4.91 751.61±21.6 3296.86±388.05 892.18±39.2 3183.16±371.60 ND 97.00±7.07 3999.01±835.19 4674.76±69.23 6149.40±82.73 21 3-辛酮 ND 153.82±18.98 36.40±5.40 ND ND 239.16±25.07 155.18±6.95 12.67±1.91 ND ND 22 2-壬酮 ND 7.06±2.53 ND ND ND 23.28±4.59 11.06±0.26 ND ND ND 23 庚烷 ND 12.18±1.72 ND ND ND 50.29±1.05 42.88±2.75 ND ND ND 24 甲苯 ND 13.16±3.03 ND ND ND 4.76±0.37 9.24±3.06 ND ND ND 25 正辛烷 ND 62.67±15.39 ND ND 13.09±2.62 64.22±5.21 125.71±26.06 ND ND 23.76±3.66 26 2-氯辛烷 ND ND ND ND ND 106.26±5.67 ND ND ND ND 27 苯乙烯 71.36±19.52 3.77±1.03 65.57±4.48 ND ND ND ND 58.63±3.65 ND ND 28 2,4-二甲基己烷 ND ND ND 14.76±1.62 ND 23.23±4.19 4.64±1.56 ND 13.65±1.30 ND 29 乙酸苯乙酯 ND ND ND 35.61±1.18 39.23±8.05 ND ND ND ND 168.47±10.49 30 甲酸辛酯 ND ND ND ND ND 6.04±0.27 ND ND ND ND 31 己酸乙烯酯 98.58±20.77 ND ND 403.93±22.3 409.14±44.23 11.06±1.64 ND 11.06±1.64 563.31±22.84 145.41±14.46 32 氯甲酸正辛酯 3.95±0.98 13.43±1.29 ND 6.48±1.58 ND ND ND ND ND ND 33 甲酸己酯 ND 75.27±9.56 ND ND 161.69±16.26 ND ND ND ND ND 34 肼基甲酸苄酯 ND ND ND 4.97±0.95 ND ND ND ND ND 24.24±4.28 35 丙位癸内酯 ND ND ND ND ND ND ND ND 76.53±4.68 78.68±6.97 36 3-甲基戊酸 49.07±16.32 ND ND ND ND 162.37±40.68 ND ND ND ND 37 异戊酸 ND ND 10.08±1.42 ND ND 52.66±7.24 ND 155.85±22.40 ND 214.84±33.84 38 异丁酸 ND ND ND ND ND 25.32±2.19 ND ND ND ND 39 2-甲基丁酸 ND 8.52±1.32 74.41±5.58 ND ND 60.85±3.03 ND 25.72±6.38 ND ND 40 2-正戊基呋喃 ND 120.39±7.92 12.11±1.13 ND ND 133.16±7.54 123.59±28.05 ND 15.98±2.45 ND 41 2,3,5,6-四甲基吡嗪 ND ND ND ND ND ND ND ND 15.38±1.67 18.19±1.03 注:“ND”表示未检出。 -
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