生物技术通报 ›› 2023, Vol. 39 ›› Issue (4): 296-303.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0954
严涛(), 陈珂可, 杨恒飞, 朱建国, 夏九学, 方曙光()
收稿日期:
2022-07-31
出版日期:
2023-04-26
发布日期:
2023-05-16
通讯作者:
方曙光,男,博士,高级工程师,研究方向:益生菌;E-mail: frank.fang@wecare-bio.com作者简介:
严涛,男,硕士,中级工程师,研究方向:益生菌工艺;E-mail: tao.yan@wecare-bio.com
基金资助:
YAN Tao(), CHEN Ke-ke, YANG Heng-fei, ZHU Jian-guo, XIA Jiu-xue, FANG Shu-guang()
Received:
2022-07-31
Published:
2023-04-26
Online:
2023-05-16
摘要:
对乳杆菌、双歧杆菌、球菌3种类型的益生菌粉的贮存活性影响因素进行研究,为其菌粉的质量指标控制及应用提供指导。以嗜酸乳杆菌LA85、植物乳杆菌Lp90、鼠李糖乳杆菌LRa05、干酪乳杆菌LC89、乳双歧杆菌BLa80、长双歧杆菌BL21、短双歧杆菌BBr60、婴儿双歧杆菌BI45、嗜热链球菌ST81、乳酸片球菌PA53、戊糖片球菌PP06、乳酸乳球菌LLa61为研究对象,以菌体的存活率、蛋白酶活性为指标,考察益生菌菌粉质量指标(水分、水分活度)、贮存温度及封装方式对其贮存过程中菌体的存活率及蛋白酶活性的影响。菌粉的水分、水分活度在一定范围内对其储存的存活率及蛋白酶活力无显著影响(P<0.05),超过一定范围,随着水分、水分活度的提升,其菌粉的储存存活率、蛋白酶活力显著下降(P<0.05);不同的贮存温度(-18℃、4℃、25℃)对菌粉的贮存存活率、蛋白酶活性有显著性影响(P<0.05);真空封装方式菌体的存活率优于普通封装(P<0.05)。确定了乳杆菌、双歧杆菌、球菌菌粉的最佳水分、水分活度的质量控制指标、贮存温度及封装方式,保证益生菌菌粉储存过程中的存活率,为益生菌厂家的菌粉质量控制标准的制定及其应用提供指导。
严涛, 陈珂可, 杨恒飞, 朱建国, 夏九学, 方曙光. 益生菌菌粉贮存活性影响因素研究[J]. 生物技术通报, 2023, 39(4): 296-303.
YAN Tao, CHEN Ke-ke, YANG Heng-fei, ZHU Jian-guo, XIA Jiu-xue, FANG Shu-guang. Study on Factors Affecting the Storage Survival Rates of Probiotic Bacteria Powder[J]. Biotechnology Bulletin, 2023, 39(4): 296-303.
菌株Strain | 批次1 Batch 1 | 批次2 Batch 2 | 批次3 Batch 3 | 批次4 Batch 4 | 批次5 Batch 5 | ||||||
---|---|---|---|---|---|---|---|---|---|---|---|
水分Moisture/% | 水分活度Water activity(Aw) | 水分Moisture/% | 水分活度Water activity(Aw) | 水分Moisture/% | 水分活度Water activity(Aw) | 水分Moisture/% | 水分活度Water activity(Aw) | 水分 Moisture/% | 水分活度Water activity(Aw) | ||
乳杆菌类Lactobacillus | LA85 | 2.70±0.21 | 0.061±0.004 | 3.57±0.22 | 0.093±0.005 | 5.18±0.23 | 0.123±0.006 | 5.68±0.25 | 0.228±0.007 | 6.78±0.26 | 0.292±0.008 |
Lp90 | 2.97±0.11 | 0.059±0.001 | 3.60±0.12 | 0.078±0.002 | 5.20±0.13 | 0.128±0.003 | 5.80±0.14 | 0.152±0.004 | 6.61±0.15 | 0.180±0.005 | |
LRa05 | 2.78±0.09 | 0.064±0.003 | 3.86±0.11 | 0.099±0.004 | 5.58±0.13 | 0.131±0.005 | 6.78±0.15 | 0.151±0.006 | 7.04±0.16 | 0.251±0.007 | |
LC89 | 3.08±0.10 | 0.060±0.004 | 3.44±0.12 | 0.068±0.005 | 5.16±0.13 | 0.126±0.006 | 5.51±0.15 | 0.154±0.007 | 6.56±0.17 | 0.175±0.008 | |
双歧杆菌类Bifidobacterium | BLa80 | 3.26±0.13 | 0.057±0.012 | 4.86±0.15 | 0.079±0.013 | 5.00±0.16 | 0.120±0.014 | 5.58±0.17 | 0.140±0.015 | 6.27±0.18 | 0.160±0.016 |
BL21 | 3.08±0.11 | 0.064±0.008 | 4.65±0.12 | 0.089±0.009 | 5.01±0.13 | 0.121±0.010 | 5.58±0.14 | 0.138±0.011 | 6.86±0.15 | 0.148±0.012 | |
BBr60 | 3.04±0.16 | 0.067±0.017 | 4.31±0.18 | 0.091±0.018 | 5.08±0.19 | 0.125±0.019 | 5.59±0.20 | 0.130±0.020 | 6.79±0.22 | 0.145±0.022 | |
BI45 | 3.12±0.17 | 0.070±0.003 | 4.31±0.19 | 0.098±0.005 | 5.07±0.20 | 0.128±0.005 | 5.55±0.21 | 0.131±0.006 | 6.89±0.25 | 0.149±0.007 | |
球菌类Cocci | ST81 | 2.52±0.08 | 0.051±0.004 | 3.07±0.09 | 0.085±0.005 | 5.15±0.10 | 0.127±0.006 | 5.55±0.11 | 0.134±0.007 | 6.48±0.13 | 0.143±0.008 |
PA53 | 2.90±0.09 | 0.058±0.011 | 3.65±0.10 | 0.088±0.012 | 5.18±0.11 | 0.127±0.013 | 5.56±0.12 | 0.138±0.014 | 6.79±0.14 | 0.141±0.015 | |
PP06 | 2.95±0.18 | 0.070±0.002 | 3.78±0.19 | 0.090±0.003 | 5.38±0.20 | 0.128±0.004 | 5.66±0.21 | 0.130±0.005 | 6.81±0.23 | 0.136±0.006 | |
LLa61 | 2.62±0.16 | 0.063±0.003 | 3.08±0.18 | 0.089±0.004 | 5.20±0.19 | 0.129±0.005 | 5.79±0.21 | 0.134±0.006 | 6.30±0.23 | 0.139±0.007 |
表1 不同水分、水分活度的益生菌菌粉
Table 1 Probiotic bacteria powder with different moisture and water activity
菌株Strain | 批次1 Batch 1 | 批次2 Batch 2 | 批次3 Batch 3 | 批次4 Batch 4 | 批次5 Batch 5 | ||||||
---|---|---|---|---|---|---|---|---|---|---|---|
水分Moisture/% | 水分活度Water activity(Aw) | 水分Moisture/% | 水分活度Water activity(Aw) | 水分Moisture/% | 水分活度Water activity(Aw) | 水分Moisture/% | 水分活度Water activity(Aw) | 水分 Moisture/% | 水分活度Water activity(Aw) | ||
乳杆菌类Lactobacillus | LA85 | 2.70±0.21 | 0.061±0.004 | 3.57±0.22 | 0.093±0.005 | 5.18±0.23 | 0.123±0.006 | 5.68±0.25 | 0.228±0.007 | 6.78±0.26 | 0.292±0.008 |
Lp90 | 2.97±0.11 | 0.059±0.001 | 3.60±0.12 | 0.078±0.002 | 5.20±0.13 | 0.128±0.003 | 5.80±0.14 | 0.152±0.004 | 6.61±0.15 | 0.180±0.005 | |
LRa05 | 2.78±0.09 | 0.064±0.003 | 3.86±0.11 | 0.099±0.004 | 5.58±0.13 | 0.131±0.005 | 6.78±0.15 | 0.151±0.006 | 7.04±0.16 | 0.251±0.007 | |
LC89 | 3.08±0.10 | 0.060±0.004 | 3.44±0.12 | 0.068±0.005 | 5.16±0.13 | 0.126±0.006 | 5.51±0.15 | 0.154±0.007 | 6.56±0.17 | 0.175±0.008 | |
双歧杆菌类Bifidobacterium | BLa80 | 3.26±0.13 | 0.057±0.012 | 4.86±0.15 | 0.079±0.013 | 5.00±0.16 | 0.120±0.014 | 5.58±0.17 | 0.140±0.015 | 6.27±0.18 | 0.160±0.016 |
BL21 | 3.08±0.11 | 0.064±0.008 | 4.65±0.12 | 0.089±0.009 | 5.01±0.13 | 0.121±0.010 | 5.58±0.14 | 0.138±0.011 | 6.86±0.15 | 0.148±0.012 | |
BBr60 | 3.04±0.16 | 0.067±0.017 | 4.31±0.18 | 0.091±0.018 | 5.08±0.19 | 0.125±0.019 | 5.59±0.20 | 0.130±0.020 | 6.79±0.22 | 0.145±0.022 | |
BI45 | 3.12±0.17 | 0.070±0.003 | 4.31±0.19 | 0.098±0.005 | 5.07±0.20 | 0.128±0.005 | 5.55±0.21 | 0.131±0.006 | 6.89±0.25 | 0.149±0.007 | |
球菌类Cocci | ST81 | 2.52±0.08 | 0.051±0.004 | 3.07±0.09 | 0.085±0.005 | 5.15±0.10 | 0.127±0.006 | 5.55±0.11 | 0.134±0.007 | 6.48±0.13 | 0.143±0.008 |
PA53 | 2.90±0.09 | 0.058±0.011 | 3.65±0.10 | 0.088±0.012 | 5.18±0.11 | 0.127±0.013 | 5.56±0.12 | 0.138±0.014 | 6.79±0.14 | 0.141±0.015 | |
PP06 | 2.95±0.18 | 0.070±0.002 | 3.78±0.19 | 0.090±0.003 | 5.38±0.20 | 0.128±0.004 | 5.66±0.21 | 0.130±0.005 | 6.81±0.23 | 0.136±0.006 | |
LLa61 | 2.62±0.16 | 0.063±0.003 | 3.08±0.18 | 0.089±0.004 | 5.20±0.19 | 0.129±0.005 | 5.79±0.21 | 0.134±0.006 | 6.30±0.23 | 0.139±0.007 |
分类Classification | 菌株Strain | 存活率Survival rate/% | |||||
---|---|---|---|---|---|---|---|
批次1 Batch 1 | 批次2 Batch 2 | 批次3 Batch 3 | 批次4 Batch 4 | 批次5 Batch 5 | |||
乳杆菌Lactobacillus | LA85 | 93.43±1.43 | 92.23±2.11 | 91.54±1.91 | 85.23±1.31* | 80.43±1.67** | |
Lp90 | 96.32±1.67 | 95.89±2.01 | 94.43±1.54 | 88.32±1.54* | 83.32±1.34** | ||
LRa05 | 95.67±1.21 | 94.65±1.81 | 93.87±1.44 | 86.32±1.67* | 81.43±2.34** | ||
LC89 | 92.54±1.67 | 92.34±1.56 | 91.67±1.89 | 83.44±1.32* | 77.34±1.43** | ||
双歧杆菌Bifidobacterium | BLa80 | 97.45±1.35 | 96.41±1.55 | 95.32±1.68 | 90.43±1.11* | 84.32±1.11** | |
BL21 | 96.65±1.22 | 95.12±1.67 | 95.31±1.44 | 90.32±1.56* | 83.21±1.51** | ||
BBr60 | 95.44±1.57 | 94.11±1.35 | 93.32±1.68 | 88.32±1.56* | 82.77±1.88** | ||
BI45 | 93.14±1.01 | 92.35±1.45 | 92.88±1.54 | 86.32±1.58* | 80.55±1.22** | ||
球菌 Cocci | ST81 | 92.55±1.65 | 91.78±1.23 | 91.11±1.63 | 85.34±1.51* | 80.69±1.51** | |
PA53 | 92.67±1.64 | 91.77±1.88 | 90.88±1.13 | 85.99±1.99* | 80.01±1.67** | ||
PP06 | 92.33±1.54 | 92.11±1.33 | 91.76±2.13 | 86.11±1.78* | 81.35±1.32** | ||
LLa61 | 88.18±1.48 | 87.44±1.54 | 86.89±1.56 | 80.23±1.65* | 74.67±1.78** |
表2 不同水分、水分活度的益生菌菌粉贮存存活率
Table 2 Storage survival rate of probiotic bacteria powder with different moisture and water activity
分类Classification | 菌株Strain | 存活率Survival rate/% | |||||
---|---|---|---|---|---|---|---|
批次1 Batch 1 | 批次2 Batch 2 | 批次3 Batch 3 | 批次4 Batch 4 | 批次5 Batch 5 | |||
乳杆菌Lactobacillus | LA85 | 93.43±1.43 | 92.23±2.11 | 91.54±1.91 | 85.23±1.31* | 80.43±1.67** | |
Lp90 | 96.32±1.67 | 95.89±2.01 | 94.43±1.54 | 88.32±1.54* | 83.32±1.34** | ||
LRa05 | 95.67±1.21 | 94.65±1.81 | 93.87±1.44 | 86.32±1.67* | 81.43±2.34** | ||
LC89 | 92.54±1.67 | 92.34±1.56 | 91.67±1.89 | 83.44±1.32* | 77.34±1.43** | ||
双歧杆菌Bifidobacterium | BLa80 | 97.45±1.35 | 96.41±1.55 | 95.32±1.68 | 90.43±1.11* | 84.32±1.11** | |
BL21 | 96.65±1.22 | 95.12±1.67 | 95.31±1.44 | 90.32±1.56* | 83.21±1.51** | ||
BBr60 | 95.44±1.57 | 94.11±1.35 | 93.32±1.68 | 88.32±1.56* | 82.77±1.88** | ||
BI45 | 93.14±1.01 | 92.35±1.45 | 92.88±1.54 | 86.32±1.58* | 80.55±1.22** | ||
球菌 Cocci | ST81 | 92.55±1.65 | 91.78±1.23 | 91.11±1.63 | 85.34±1.51* | 80.69±1.51** | |
PA53 | 92.67±1.64 | 91.77±1.88 | 90.88±1.13 | 85.99±1.99* | 80.01±1.67** | ||
PP06 | 92.33±1.54 | 92.11±1.33 | 91.76±2.13 | 86.11±1.78* | 81.35±1.32** | ||
LLa61 | 88.18±1.48 | 87.44±1.54 | 86.89±1.56 | 80.23±1.65* | 74.67±1.78** |
分类Classification | 菌株Strain | 存活率Survival rate/% | |||
---|---|---|---|---|---|
贮存-18℃Storage at -18℃ | 贮存4℃Storage at 4℃ | 贮存25℃Storage at 25℃ | |||
乳杆菌 Lactobacillus | LA85 | 92.23±2.11 | 79.34±1.91** | 48.56±2.67** | |
Lp90 | 95.12±1.81 | 84.34±1.22** | 44.31±1.79** | ||
LRa05 | 93.33±1.22 | 83.23±1.87** | 40.36±1.64** | ||
LC89 | 90.01±1.12 | 82.15±1.85** | 43.43±1.57** | ||
双歧杆菌 Bifidobacterium | BLa80 | 94.45±1.32 | 84.54±1.67** | 49.33±1.67** | |
BL21 | 92.65±1.32 | 78.24±1.86** | 31.35±1.65** | ||
BBr60 | 92.67±1.33 | 82.34±1.66** | 36.34±2.66** | ||
BI45 | 94.54±1.45 | 79.11±1.89** | 26.24±1.76** | ||
球菌 Cocci | ST81 | 90.45±1.55 | 72.11±2.89** | 43.33±1.67** | |
PA53 | 91.20±1.01 | 72.45±1.55** | 33.34±1.66** | ||
PP06 | 90.03±1.06 | 78.13±1.97** | 45.33±1.67** | ||
LLa61 | 86.78±1.22 | 71.43±1.57** | 43.53±2.47** |
表3 不同贮存温度对益生菌菌粉贮存活性影响
Table 3 Effects of different storage temperatures on the storage survival rates of probiotic bacteria powder
分类Classification | 菌株Strain | 存活率Survival rate/% | |||
---|---|---|---|---|---|
贮存-18℃Storage at -18℃ | 贮存4℃Storage at 4℃ | 贮存25℃Storage at 25℃ | |||
乳杆菌 Lactobacillus | LA85 | 92.23±2.11 | 79.34±1.91** | 48.56±2.67** | |
Lp90 | 95.12±1.81 | 84.34±1.22** | 44.31±1.79** | ||
LRa05 | 93.33±1.22 | 83.23±1.87** | 40.36±1.64** | ||
LC89 | 90.01±1.12 | 82.15±1.85** | 43.43±1.57** | ||
双歧杆菌 Bifidobacterium | BLa80 | 94.45±1.32 | 84.54±1.67** | 49.33±1.67** | |
BL21 | 92.65±1.32 | 78.24±1.86** | 31.35±1.65** | ||
BBr60 | 92.67±1.33 | 82.34±1.66** | 36.34±2.66** | ||
BI45 | 94.54±1.45 | 79.11±1.89** | 26.24±1.76** | ||
球菌 Cocci | ST81 | 90.45±1.55 | 72.11±2.89** | 43.33±1.67** | |
PA53 | 91.20±1.01 | 72.45±1.55** | 33.34±1.66** | ||
PP06 | 90.03±1.06 | 78.13±1.97** | 45.33±1.67** | ||
LLa61 | 86.78±1.22 | 71.43±1.57** | 43.53±2.47** |
分类Classification | 菌株Strain | 存活率Survival rate/% | |||
---|---|---|---|---|---|
普通封装Ordinary encapsulation | 真空封装Vacuum encapsulation | 氮气封装Nitrogen encapsulation | |||
乳杆菌 Lactobacillus | LA85 | 91.35±1.81 | 95.01±1.01* | 94.56±1.44* | |
Lp90 | 93.22±1.81 | 97.12±1.22* | 96.65±1.78* | ||
LRa05 | 91.11±1.81 | 96.13±1.55* | 96.01±1.32* | ||
LC89 | 89.78±2.11 | 94.32±1.32* | 93.68±1.33* | ||
双歧杆菌Bifidobacterium | BLa80 | 91.67±1.33 | 96.23±1.22* | 96.15±1.22* | |
BL21 | 90.78±1.22 | 95.32±1.68* | 95.32±1.45* | ||
BBr60 | 91.67±1.34 | 95.89±1.01* | 95.22±1.11* | ||
BI45 | 92.32±1.68 | 96.55±1.32* | 95.82±1.22* | ||
球菌 Cocci | ST81 | 88.67±1.31 | 93.89±1.11* | 93.43±1.42* | |
PA53 | 89.89±1.11 | 94.89±1.56* | 94.32±1.66* | ||
PP06 | 88.11±1.89 | 93.65±1.34* | 93.01±1.32* | ||
LLa61 | 84.89±1.14 | 89.43±1.32* | 89.21±1.21* |
表4 不同包装方式对益生菌菌粉贮存活性影响
Table 4 Effects of different packaging methods on the storage survival rates of probiotic bacteria powder
分类Classification | 菌株Strain | 存活率Survival rate/% | |||
---|---|---|---|---|---|
普通封装Ordinary encapsulation | 真空封装Vacuum encapsulation | 氮气封装Nitrogen encapsulation | |||
乳杆菌 Lactobacillus | LA85 | 91.35±1.81 | 95.01±1.01* | 94.56±1.44* | |
Lp90 | 93.22±1.81 | 97.12±1.22* | 96.65±1.78* | ||
LRa05 | 91.11±1.81 | 96.13±1.55* | 96.01±1.32* | ||
LC89 | 89.78±2.11 | 94.32±1.32* | 93.68±1.33* | ||
双歧杆菌Bifidobacterium | BLa80 | 91.67±1.33 | 96.23±1.22* | 96.15±1.22* | |
BL21 | 90.78±1.22 | 95.32±1.68* | 95.32±1.45* | ||
BBr60 | 91.67±1.34 | 95.89±1.01* | 95.22±1.11* | ||
BI45 | 92.32±1.68 | 96.55±1.32* | 95.82±1.22* | ||
球菌 Cocci | ST81 | 88.67±1.31 | 93.89±1.11* | 93.43±1.42* | |
PA53 | 89.89±1.11 | 94.89±1.56* | 94.32±1.66* | ||
PP06 | 88.11±1.89 | 93.65±1.34* | 93.01±1.32* | ||
LLa61 | 84.89±1.14 | 89.43±1.32* | 89.21±1.21* |
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