纺织学报 ›› 2022, Vol. 43 ›› Issue (12): 82-87.doi: 10.13475/j.fzxb.20211108006

• 染整与化学品 • 上一篇    下一篇

罗布麻微生物脱胶的菌种筛选与工艺优化

程芬1, 张兴群1,2, 王云龙1, 王颖1()   

  1. 1.新疆大学 纺织与服装学院, 新疆 乌鲁木齐 830046
    2.东华大学 生物与医学工程学院, 上海 201620
  • 收稿日期:2021-11-17 修回日期:2022-07-23 出版日期:2022-12-15 发布日期:2023-01-06
  • 通讯作者: 王颖
  • 作者简介:程芬(1995—),女,硕士生。主要研究方向为纺织生物技术与生物医用材料。
  • 基金资助:
    国家自然科学基金项目(51863020);天池博士计划项目(042312060);新疆大学博士科研启动基金项目(2019720001)

Optimizing microorganism degumming process of apocynum and bacterial screening

CHENG Fen1, ZHANG Xingqun1,2, WANG Yunlong1, WANG Ying1()   

  1. 1. College of Textile and Clothing, Xinjiang University, Urumqi, Xinjiang 830046, China
    2. College of Biological Sciences and Medical Engineering, Donghua University, Shanghai 201620, China
  • Received:2021-11-17 Revised:2022-07-23 Published:2022-12-15 Online:2023-01-06
  • Contact: WANG Ying

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摘要：

为提高罗布麻微生物脱胶效率,采用透明圈法、DNS比色法以及16S rRNA分子鉴定等方法从新疆乌鲁木齐南山野生麻生长区土壤中筛选优势脱胶菌株,并设计正交试验将所筛优势菌株用于优化新疆罗布麻微生物脱胶工艺参数。试验结果表明:所筛7株菌具有较高果胶酶活、木聚糖酶活,低纤维素酶活,适用于罗布麻微生物脱胶的实际应用;7菌种主要为芽孢杆菌属,此外还有肠杆菌属、克雷伯菌属和泛菌属;在脱胶液pH值为7,浴比为1∶40,摇床转速为90 r/min的最优脱胶工艺参数下,最佳脱胶方式为7菌系复合脱胶,脱胶时残胶率可降至30.45%,脱胶效果改善,同时菌株之间形成稳定的脱胶菌群,脱胶时菌种之间的协同作用、拮抗作用会影响菌群的脱胶能力。

关键词: 罗布麻, 菌种筛选, 酶活, 微生物脱胶, 脱胶工艺

Abstract:

In order to improve the efficiency of apocynum degumming, dominant degumming strains was screened from the soil of the wild hemp growth area of Urumqi, Xinjiang using the transparent zones method, DNS colorimetry method and 16S rRNA sequencing. The superior degumming strains were used to optimize the microbial degumming process of apocynum by the orthogonal experiment designed in this study. The experimental results show that seven dominant degumming strains with higher pectinase, xylanase and low cellulase are theoretically suitable for practical applications of apocynum degumming, which belong to four genera including Bacillus, Enterobacter, Klebsiella and Pantoea. The optimal parameters optimized by orthogonal tests are initial pH 7 of the degumming solution, bath ratio of 1:40 and shaker speed of 90 r/min. Using the optimization process parameters, the residual gum rate of complex bacteria can be reduced to 30.45%, the degumming effect is improved and thus the best method is degumming with 7 strains, because a stable degumming colony is formed between strains among which the synergism and antagonism may affect the degumming ability.

Key words: apocynum, bacterial screening, enzyme activity, microbial degumming, degumming process

中图分类号: 

  • TS192.5

表1

糖标液配制"

试管号 糖标液体积 去离子水体积 DNS体积
1 0.0 2.0 2.0
2 0.2 1.8 2.0
3 0.4 1.6 2.0
4 0.5 1.5 2.0
5 0.6 1.4 2.0
6 0.7 1.3 2.0
7 0.8 1.2 2.0
8 1.0 1.0 2.0

表2

正交试验因素水平表"

水平 A
摇床转速/(r·min-1)		 B
pH值		 C
浴比
1 0 7 1∶30
2 90 8 1∶40
3 150 9 1∶50

图1

菌株刚果红HC值"

表3

酶活测定结果"

菌株编号 果胶酶活 木聚糖酶活 纤维素酶酶活
Y8 117.565 9.390 2.825
X7 54.426 33.470 0.220
YE13 83.217 66.470 -
LT3 62.507 28.110 -
Z4 139.032 31.280 -
2-1 141.557 - -
X5 61.118 55.560 -
L1 95.340 50.600 -
GAN1 106.578 104.890 -

表4

菌株基因序列比对结果"

菌株编号 细菌种属 GenBank登录号 相似性/%
2-1 芽孢杆菌属
(Bacillus subtilis subsp.)		 NR 112686.1 100.00
X7 克雷伯菌属
(Klebsiella sp.)		 MK 780166.1 100.00
YE13 芽孢杆菌属
(Bacillus subtilis subsp.)		 MK 875169.1 100.00
L1 大肠杆菌属
(Enterobacter sp.)		 MK 318778.1 99.36
GAN1 泛菌属
(Pantoea sp.)		 FJ 357809.7 98.73
Y8 芽孢杆菌属
(Bacillus subtilis subsp.)		 MN 744459.1 100.00
Z4 环状芽孢杆菌属
(Bacillus circulans strain)		 CP 026033.1 99.86

表5

微生物脱胶正交试验优化结果"

试验
号		 A B C 空列D 残胶
率/%		 断裂强度/
(cN·dtex-1)
1 1 1 1 1 47.76 6.33
2 1 2 2 2 44.69 1.82
3 1 3 3 3 42.90 3.13
4 2 1 2 3 36.86 8.19
5 2 2 3 1 42.52 4.28
6 2 3 1 2 41.28 3.08
7 3 1 3 2 40.22 2.22
8 3 2 1 3 46.64 2.56
9 3 3 2 1 38.97 1.72
残胶率 K1 45.12 41.61 45.23 43.08
K2 40.22 44.62 40.17 42.06
K3 41.94 41.05 41.88 42.13
R 4.90 3.57 5.05 1.02
断裂强度 K1 3.76 5.58 3.99 4.11
K2 5.18 2.89 3.91 2.37
K3 2.17 2.64 3.21 4.63
R' 3.01 2.94 0.78 2.25

表6

方差分析表"

指标 方差来源 平方和 自由度 均方和 F比 显著性
残胶率 A 37.08 2 18.54 19.04 **
B 22.12 2 11.06 11.36 *
C 39.75 2 19.88 20.41 **
D 1.95 2 0.97
断裂
强度		 A 13.60 2 6.80 12.31 *
B 15.94 2 7.97 14.43 *
C 1.10 2 0.55
D 8.41 2 4.20 7.61

表7

脱胶效果验证表"

菌株 残胶
率/%		 菌株 残胶
率/%
Z4 57.70 C6/1(Z4,Y8,GAN1,2-1,L1,X7) 50.74
YE13 57.00 C6/2(Z4,YE13,Y8,2-1,L1,X7) 44.06
Y8 55.50 C6/3(YE13,Y8,GAN1,2-1,L1,X7) 42.86
GAN1 50.90 C6/4(Z4,YE13,GAN1,2-1,L1,X7) 38.60
2-1 48.40 C6/5(Z4,YE13,Y8,GAN1,2-1,X7) 34.70
L1 47.70 C6/6(Z4,YE13,Y8,GAN1,2-1,L1) 32.19
X7 46.64 C6/7(Z4,YE13,Y8,GAN1,L1,X7) 31.04
C7(Z4,YE13,Y8,GAN1,2-1,L1,X7) 30.45
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