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饲草附生乳酸菌对碳源的选择性

陈鑫珠 董朝霞 张建国

陈鑫珠,董朝霞,张建国. 饲草附生乳酸菌对碳源的选择性 [J]. 福建农业学报,2024,39(5):1−10
引用本文: 陈鑫珠,董朝霞,张建国. 饲草附生乳酸菌对碳源的选择性 [J]. 福建农业学报,2024,39(5):1−10
CHEN X Z, DONG Z X, ZHANG J G. Silage Carbon Sources Preferred by Epiphytic Lactic Acid Bacteria [J]. Fujian Journal of Agricultural Sciences,2024,39(5):1−10
Citation: CHEN X Z, DONG Z X, ZHANG J G. Silage Carbon Sources Preferred by Epiphytic Lactic Acid Bacteria [J]. Fujian Journal of Agricultural Sciences,2024,39(5):1−10

饲草附生乳酸菌对碳源的选择性

基金项目: 国家自然科学基金项目(31971764、32302791);福建省农业高质量发展超越“5511”协同创新工程项目(XTCXGC2021019);福建省科技计划公益类专项(2020R1021002)
详细信息
    作者简介:

    陈鑫珠(1985 —),女,博士,副研究员,主要从事动物营养、非常规饲料资源的开发利用和饲草加工与贮藏研究, E-mail:010622051@163.com

    通讯作者:

    张建国(1968 —),男,博士生导师,教授,主要从事饲草料加工利用、生物活性成分研究,E-mail:zhangjg@scau.edu.cn

  • 中图分类号: S816.53

Silage Carbon Sources Preferred by Epiphytic Lactic Acid Bacteria

  • 摘要:   目的  探究饲草源乳酸菌对碳源的选择性,为不同草种筛选适宜的青贮菌种。  方法  选取饲草中分离获得的16株代表性乳酸菌作为研究对象,在MRS液体培养基中以葡萄糖、蔗糖、果糖、木糖、棉籽糖以及5种糖混合为碳源,进行不同菌株发酵培养。每株乳酸菌设3个重复,每个糖处理设空白对照(不接菌)。采用高效液相色谱仪测定各处理发酵24 h后液体培养基中剩余碳源含量和乳酸产量。  结果  (1)蔗糖是大部分菌株发酵首选碳源,在棉籽糖、葡萄糖、果糖和木糖的选择上不同菌株差异较大,发酵乳杆菌(Lactobacillus fermentum,LbF-WM)首选棉籽糖和木糖,假肠膜明串珠菌(Leuconostioc pseudomesenteroides,LeP-IR)首选木糖。(2)罗伊氏乳杆菌(Lactobacillus reuteri,LbR-EG)和柠檬明串珠菌(Leuconostoc citreum,LeC-IR)对蔗糖,LbF-WM和LbR-EG对果糖, LbF-WM 、LbR-EG和LeP-IR对棉籽糖的利用率均达100%,屎肠球菌(Enterococcus faecium,EF-KG)、短乳杆菌(Lactobacillus brevis,LbB-KG)、戊糖乳杆菌(Lactobacillus pentosus,LbPe-EG)、乳酸乳球菌(Lactococcus lactis,LcL-SC)对葡萄糖的利用率高于其他碳源。(3)灰黄色肠球菌(Enterococcus durans,EG-IR)利用葡萄糖和混合糖,LbPe-EG利用果糖,肠膜明串珠菌(Leuconostoc mesenteroides,LeM-IR)利用棉籽糖和混合糖,LbPe-EG利用蔗糖,LbF-WM利用木糖,格氏乳球菌(Lactococcus garvieae,LcG-IR)利用混合糖发酵乳酸产量最高。(4)耐久肠球菌(Enterococcus durans,ED-IR)利用葡萄糖,植物乳杆菌(Lactobacillus plantarum,LbPl-IR)、LbPe-EG利用果糖, LbPl-IR和LeM-IR利用棉籽糖, LbPe-EG、LbPl-IR和ED-IR利用蔗糖, LeP- IR利用木糖,LcG-IR和EG-IR利用混合糖产乳酸效率较高。LbPl-IR在5种单糖中产酸效率均达到80%以上。  结论  不同菌株对同一碳源的利用率、产酸量和产酸效率均不同;同一株菌株对不同碳源的利用率、产酸量和产酸效率也不相同。其中,蔗糖为大部分菌株的首选碳源,蔗糖和葡萄糖的利用率和产乳酸量较高,葡萄糖的产酸效率较高;青贮调制时,可针对饲草中分布乳酸菌种类选择最适碳源。
  • 图  1  16株乳酸菌培养24 h后在5种糖混合培养基中各种糖的剩余含量

    1)图中柱状数据表示混合糖培养基中剩余各糖的百分含量。2)CK,无接菌的混合糖培养基;

    Figure  1.  Contents of residual sugars in mixed sugar media of 16 different LAB cultured for 24 h

    1) Data on a column are contents of residual individual sugars in medium of mixed sugars; 2) CK without inoculation on medium of mixed sugars; L.M.: Le. mesenteroides; L.P.: Le. pseudomesenteroides; W.P.: W. paramesenteroides. Same for below.

    表  1  16株乳酸菌的名称及来源

    Table  1.   Names and sources of 16 LAB strains

    序号
    No.
    命名
    Name
    种名
    Species
    来源
    Source
    序号
    No.
    命名
    Name
    种名
    Species
    来源
    Source
    1 EF-KG 屎肠球菌 E. faecium 热研四号王草 9 LcG-IR 格氏乳球菌 Lc. garvieae 意大利黑麦草
    2 ED-IR 耐久肠球菌 E. durans 意大利黑麦草 10 LcL-SC 乳酸乳球菌 Lc. lactis 甜玉米
    3 EG-IR 灰黄色肠球菌 E. gilvus 意大利黑麦草 11 LeC-IR 柠檬明串珠菌 Le. citreum 意大利黑麦草
    4 LbB-KG 短乳杆菌 Lb. brevis 热研四号王草 12 LeM-IR 肠膜明串珠菌 Le. mesenteroides 意大利黑麦草
    5 LbF-WM 发酵乳杆菌 Lb. fermentum 糯玉米 13 LeP-IR 假肠膜明串珠菌 Le. pseudomesenteroides 意大利黑麦草
    6 LbPe-EG 戊糖乳杆菌 Lb. pentosus 摩特矮象草 14 WCo-KG 融合乳杆菌 W. confusa 热研四号王草
    7 LbPl-IR 植物乳杆菌 Lb. plantarum 意大利黑麦草 15 WCi-MT 魏斯特菌 W. cibaria MT-1杂交象草
    8 LbR-EG 罗伊氏乳杆菌 Lb. reuteri 摩特矮象草 16 WP-IR 类肠膜魏斯氏菌 W. paramesenteroides 意大利黑麦草
    下载: 导出CSV

    表  2  牧草原料中糖分组成

    Table  2.   Sugars in native grass material (%)

    项目
    Item
    葡萄糖
    Glucose
    蔗糖
    Sucrose
    果糖
    Fructose
    棉籽糖
    Raffinose
    木糖
    Xylose
    热研四号王草
    Pennisetum purpureum × P. americanum cv. Reyan No.4
    0.07±0.01c 1.91±0.37a 1.34±0.23b 1.86±0.06a 0.00±0.00c
    MT-1象草
    P. purpureum cv. MT-1
    0.10±0.01c 1.82±0.08a 1.41±0.25b 1.28±0.27b 0.03±0.00c
    摩特矮象草
    P. purputeum cv. Mott
    0.12±0.01c 1.75±0.29ab 2.11±0.13a 1.58±0.31b 0.09±0.09c
    甜玉米
    Zea mays L. saccharata Sturt.
    1.84±0.26b 6.00±0.96a 1.12±0.11b 0.00±0.00c 0.00±0.00c
    桂牧一号杂交象草
    (P. americanum × P. Purpureum) × (P. Purpureum cv. Mott) Guimu No.1
    0.03±0.02d 1.87±0.14a 0.21±0.07c 1.24±0.17b 0.00±0.00d
    糯玉米
    Zea mays L. Sinensisa Kulesh
    1.81±0.36b 5.06±0.37a 0.93±0.02c 0.00±0.00d 0.00±0.00d
    意大利黑麦草
    Lolium multiflorum
    0.70±0.39ab 0.94±0.36a 0.34±0.07b 0.08±0.08c 0.10±0.04c
    同行数据后不同小写字母表示差异显著(P<0.05),相同或无字母表示差异不显著(P>0.05);下表同。
    Data with different lowercase letters on same row represent significant difference at P<0.05; those with same or no letter, no significant difference at P>0.05. Same for below.
    下载: 导出CSV

    表  3  16株乳酸菌对6种糖源培养液发酵后的碳源利用率

    Table  3.   Sugar utilization by 16 different LAB in 6 treatments (单位:%)

    菌株编号
    Strain code
    葡萄糖MRS培养基
    MRS-P
    果糖MRS培养基
    MRS-G
    棉籽糖MRS培养基
    MRS-MZ
    蔗糖MRS培养基
    MRS-Z
    木糖MRS培养基
    MRS-M
    混合糖MRS培养基
    MRS- MIX
    EF-KG 94.68±1.01a 29.25±3.43e 0.73±0.53f 89.99±1.72b 59.95±1.48c 50.27±0.54d
    ED-IR 44.76±2.10b 60.48±1.52a 63.67±0.91a 69.87±1.16a 5.22±0.18c 39.83±2.34b
    EG-IR 71.44±3.35ab 58.97±3.27bc 49.20±1.22cd 81.23±1.54a 0.00±0.00e 39.56±0.89d
    LbB-KG 72.13±1.05a 18.10±0.53c 7.80±4.49c 50.41±0.10b 46.74±0.10b 38.82±2.37b
    LbF-WM 87.50±1.43b 100.00±0.00a 100.00±0.00a 89.79±3.33b 79.15±0.84b 33.33±4.13c
    LbPe-EG 82.97±0.73a 60.31±0.62b 37.42±3.02c 76.75±1.05ab 1.10±0.19d 38.56±2.88c
    LbPl-IR 69.56±1.06a 58.45±1.72a 60.34±1.81a 72.80±2.56a 0c 39.57±2.86b
    LbR-EG 82.07±2.63b 100.00±0.00a 100.00±0.00a 100.00±0.00a 0d 31.01±5.99c
    LcG-IR 78.09±0.84ab 60.59±1.02b 74.69±0.71ab 81.48±1.64a 1.92±0.42d 38.11±3.78c
    LcL-SC 93.58±1.01a 0e 18.52±0.52d 90.29±0.82a 37.25±0.12c 52.81±5.62b
    LeC-IR 83.17±3.13b 79.15±0.61b 10.49±0.03d 100.00±0.00a 15.54±0.16d 44.51±10.98c
    LeM-IR 73.89±1.05a 55.36±4.22b 70.94±0.01a 74.02±0.65a 0d 24.70±4.80c
    LeP-IR 80.19±1.64b 90.59±0.10a 100.00±0.00a 52.23±6.10c 24.86±20.14c 33.81±2.39c
    WCo-KG 76.66±1.64b 9.93±5.08e 17.11±1.42d 96.40±1.00a 48.27±0.10c 53.27±6.55c
    WCi-MT 67.93±3.46a 56.89±0.82a 68.83±1.55a 71.00±0.86a 8.80±0.18b 27.70±7.52b
    WP-IR 79.14±1.24a 73.84±0.01a 79.16±2.21a 79.09±1.13a 0c 41.12±7.76b
    下载: 导出CSV

    表  4  16株乳酸菌在不同碳源培养基发酵中的乳酸产量

    Table  4.   Lactic acid production by 16 different LAB on different media (单位:%)

    菌株名称
    Name
    葡萄糖MRS培养基
    MRS-P
    果糖MRS培养基
    MRS-G
    棉籽糖MRS培养基
    MRS-MZ
    蔗糖MRS培养基
    MRS-Z
    木糖MRS培养基
    MRS-M
    混合糖MRS培养基
    MRS- MIX
    EF-KG 0.69±0.01a 0.32±0.02b 0. 01±0.00c 0.62±0.01a 0.58±0.03a 0.55±0.02a
    ED-IR 0.89±0.02a 0.85±0.05a 0.21±0.03b 1.14±0.01a 0.02±0.00c 0.86±0.06a
    EG-IR 1.25±0.05a 0.73±0.05b 0.66±0.03b 0.87±0.00b 0.01±0.00c 1.00±0.03a
    LbB-KG 0.71±0.00a 0.29±0.03c 0.23±0.00c 0.56±0.06b 0.63±0.04b 0.61±0.04b
    LbF-WM 0.67±0.01b 0.36±0.04c 0.71±0.01b 0.73±0.00b 0.94±0.00a 0.67±0.05b
    LbPe-EG 1.08±0.04b 1.10±0.06a 0.43±0.04c 1.31±0.00a 0.01±0.00c 0.98±0.02b
    LbPl-IR 1.14±0.01a 1.07±0.05a 1.01±0.02a 1.23±0.03a 0.61±0.02b 0.77±0.00b
    LbR-EG 0.73±0.02ab 0.69±0.04ab 0.86±0.02a 0.55±0.03b 0.63±0.04ab 0.67±0.00 ab
    LcG-IR 1.18±0.04a 0.88±0.00b 1.02±0.04a 1.15±0.01b a 0.31±0.00c 1.08±0.05a
    LcL-SC 0.69±0.02a 0.45±0.06b 0.15±0.00c 0.52±0.05ab 0.50±0.02ab 0.58±0.00a
    LeC-IR 0.71±0.02a 0.67±0.02a 0.14±0.05c 0.42±0.02b 0.15±0.00c 0.45±0.01b
    LeM-IR 1.13±0.04a 1.01±0.05a 1.16±0.00a 1.13±0.00a 0.71±0.01b 1.06±0.02a
    LeP-IR 0.79±0.00a 0.54±0.02c 0.76±0.02a 0.65±0.01bc 0.78±0.00a 0.61±0.01bc
    WCo-KG 0.60±0.02a 0.43±0.00b 0.13±0.00c 0.62±0.06a 0.55±0.02a 0.50±0.02ab
    WCi-MT 1.11±0.05a 0.54±0.00b 0.36±0.02c 0.67±0.05b 0.10±0.01d 0.69±0.02b
    WP-IR 0.58±0.00a 0.52±0.00a 0.08±0.01c 0.32±0.00b 0.64±0.02a 0.31±0.00b
    下载: 导出CSV

    表  5  16株乳酸菌的产酸效率

    Table  5.   Acid production efficiency of 16 different LAB (单位:%)

    菌株名称
    Name
    葡萄糖MRS培养基
    MRS-P
    果糖MRS培养基
    MRS-G
    棉籽糖MRS培养基
    MRS-MZ
    蔗糖MRS培养基
    MRS-Z
    木糖MRS培养基
    MRS-M
    混合糖MRS培养基
    MRS- MIX
    EF-KG 36.44±0.01c 54.69±0.06a 34.26±1.17d 34.45±0.01d 48.38±0.06b 54.71±0.59a
    ED-IR 99.41±0.22a 70.27±0.02d 16.49±0.00f 81.58±0.07b 19.16±0.66e 71.47±0.00c
    EG-IR 87.49±0.06a 61.90±0.02d 67.07±0.02c 53.55±0.03e 50.00±0.00f 82.72±0.00b
    LbB-KG 49.21±0.04b 80.12±0.15ab 81.69±0.20a 55.55±0.11b 67.40±0.15ab 49.85±0.00b
    LbF-WM 38.29±0.01d 18.00±0.00f 35.50±0.00e 40.65±0.01c 59.38±0.03a 49.12±0.00b
    LbPe-EG 65.08±0.02d 91.20±0.03a 57.46±0.03e 85.34±0.06b 23.10±3.98f 79.76±0.00c
    LbPl-IR 81.94±0.06d 91.53±0.03a 83.69±0.01c 84.70±0.45b 81.33±0.00e 63.71±0.00f
    LbR-EG 44.48±0.02c 34.50±0.00e 43.00±0.00d 27.50±0.00f 45.99±0.00b 48.56±0.00a
    LcG-IR 75.55±0.04b 72.61±0.02c 68.28±0.01e 70.57±0.03d 41.77±0.00f 87.25±0.00a
    LcL-SC 36.87±0.01d 66.18±0.00b 40.49±0.05e 28.80±0.01f 67.11±0.22a 61.45±0.00c
    LeC-IR 42.69±0.02c 42.32±0.01c 66.75±0.14a 21.00±0.00e 48.28±0.50b 40.55±0.00d
    LeM-IR 76.47±0.05d 91.22±0.03a 81.76±0.01b 76.33±0.05e 79.78±0.00c 70.38±0.00f
    LeP-IR 49.26±0.03c 29.80±0.01f 38.00±0.00e 62.23±0.12b 86.66±0.00a 46.08±0.00d
    WCo-KG 39.13±0.02d 43.10±0.00c 37.98±0.05e 32.16±0.00f 56.97±0.12a 53.50±0.00b
    WCi-MT 81.70±0.07a 47.46±0.02c 26.15±0.00d 47.18±0.04c 56.85±1.12b 47.72±0.00c
    WP-IR 36.65±0.02a 35.21±0.01a 5.05±0.00b 20.23±0.01ab 15.80±27.37ab 26.33±0.00ab
    下载: 导出CSV
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  • 收稿日期:  2023-11-28
  • 修回日期:  2024-02-18
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