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《山东大学学报(理学版)》 ›› 2020, Vol. 55 ›› Issue (7): 103-110.doi: 10.6040/j.issn.1671-9352.0.2019.667

•   • 上一篇    

Klebsiella sp. LDX1-1多糖的发酵影响因素和絮凝特征

王胜男1(),栾兴社2,3,*(),郭新娟3,肖敏4,戴美学1   

  1. 1. 山东师范大学生命科学学院, 山东 济南 250014
    2. 山东建筑大学市政与环境工程学院, 山东 济南 250101
    3. 青岛耀东生物工程有限公司, 山东 青岛 266109
    4. 山东大学微生物技术国家重点实验室, 国家糖工程技术研究中心, 山东 青岛 266237
  • 收稿日期:2019-09-22 出版日期:2020-07-20 发布日期:2020-07-08
  • 通讯作者: 栾兴社 E-mail:1062825736@qq.com;xingsheluan@126.com
  • 作者简介:王胜男(1996—),女,硕士研究生,研究方向为微生物多糖. E-mail:1062825736@qq.com

Influencing factors of fermentation and flocculation characteristics of Klebsiella sp. LDX1-1 polysaccharide

Sheng-nan WANG1(),Xing-she LUAN2,3,*(),Xin-juan GUO3,Min XIAO4,Mei-xue DAI1   

  1. 1. School of Life Science, Shandong Normal University, Jinan 250014, Shandong, China
    2. School of Municipal and Environmental Engineering, Shandong Jianzhu University, Jinan 250101, Shandong, China
    3. Qingdao Yaodong Bioengineering Co. Ltd, Qingdao 266109, Shandong, China
    4. State Key Laboratory of Microbial Technology and National Glycoengineering Research Center, Shandong University, Qingdao 266237, Shandong, China
  • Received:2019-09-22 Online:2020-07-20 Published:2020-07-08
  • Contact: Xing-she LUAN E-mail:1062825736@qq.com;xingsheluan@126.com

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摘要：

Klebsiella sp.LDX1-1多糖产生的最佳碳源为麦芽糖与低聚异麦芽糖的混合糖,其中低聚异麦芽糖质量分数为8%;最佳氮源为2g/L的NH4NO3。2g/L的雪松松针提取物、0.4%的曲拉通X-100显著促进多糖合成,产量分别提高12.46%和14.38%。用综合试验因素进行发酵,得到的最适发酵周期为24h,粗多糖产量达到23.46g/L,比发酵基础培养基条件下提高43.14%。由发酵液制取的初步纯化多糖具有高效絮凝活性,对高岭土悬液和黄河水的絮凝率分别是98.43%和96.31%。纯化样品的红外光谱定性分析表明:发酵产生的黏性物质成分为典型的生物多糖。

关键词: Klebsiella sp.LDX1-1, 多糖, 发酵, 絮凝

Abstract:

The best carbon source for the production of Klebsiella sp. LDX1-1 polysaccharide is the mixture of maltose and isomaltooligosaccharide, in which the mass fraction of isomaltooligosaccharide is 8%, and the best nitrogen source is NH4NO3 for 2g/L. 2g/L of cedar needle extract and 0.4% of Triton X-100 significantly promote the polysaccharide synthesis, and the yield increased by 12.46% and 14.38%, respectively. The optimum fermentation period is 24h, and the yield of crude polysaccharide is 23.46g/L, which is 43.14% higher than that under the basic fermentation conditions. The preliminary purified polysaccharide prepared from fermentation broth has high flocculation activity, and the flocculation rate of kaolin suspension and the Yellow River water is 98.43% and 96.31%, respectively. The qualitative analysis of the purified samples by infrared spectroscopy shows that the components of the viscous substances produced by fermentation are typical biological polysaccharides.

Key words: Klebsiella sp. LDX1-1, polysaccharide, fermentation, flocculation

中图分类号: 

  • Q93

表1

碳源对多糖产生的影响"

检测项目 蔗糖 葡萄糖 麦芽糖 乳糖 果糖 木糖 混合糖
发酵液pH 5.95 5.41 5.80 5.92 5.99 5.36 5.64
黏度(mPa·s) 316 0.00 403 258 110 0.00 423
粗多糖(g·L-1) 14.09 0.76 16.39 15.92 13.09 1.36 17.36
生物量(OD600) 1.957 0.859 2.059 1.864 2.094 0.94 2.142

表2

混合糖中低聚异麦芽糖质量分数对多糖产生的影响"

检测项目 对照 混合糖/%
2 4 6 8 10 12 14
发酵液pH 6.13 5.63 5.63 5.64 5.64 5.61 5.58 5.56
黏度(mPa·s) 410 462 467 486 488 503 508 511
粗多糖(g·L-1) 16.31 16.82 17.31 17.85 18.16 17.66 16.95 16.42
生物量(OD600) 1.679 1.703 1.708 1.711 1.937 1.860 1.869 1.899

表3

氮源对多糖产生的影响"

检测项目 NH4NO3 KNO3 NaNO3 (NH4)2SO4 (NH4)2HPO4 NH4Cl 尿素
发酵液pH 5.73 5.95 5.98 5.50 5.80 5.45 5.89
黏度(mPa·s) 455 426 415 279 253 203 143
粗多糖(g·L-1) 18.26 18.12 18.08 14.05 12.62 12.82 13.09
生物量(OD600) 2.071 1.788 1.810 1.736 1.913 1.946 1.840

表4

生长复合因子对多糖产生的影响"

检测项目 对照 牛肉膏 胰蛋白胨 鱼粉蛋白胨 酵母提取物 玉米浆 麸皮提取物 松针提取物
发酵液pH 5.47 5.61 5.58 5.58 5.71 5.28 5.44 6.33
黏度(mPa·s) 451 568 573 579 550 467 346 526
粗多糖(g/L) 18.21 19.33 19.26 19.29 18.27 18.63 18.48 20.48
生物量(OD600) 1.693 1.889 1.878 1.880 2.011 1.773 1.829 1.705

表5

曲拉通X-100对多糖产生的影响"

检测项目 对照 0.1% 0.2% 0.3% 0.4% 0.5% 0.6% 0.7%
发酵液pH 6.31 5.61 5.45 5.49 5.46 5.38 5.36 5.35
粘度(mPa·s) 523 550 578 585 589 567 546 533
粗多糖(g·L-1) 20.51 21.72 21.81 22.26 23.46 22.53 21.02 20.81
生物量(OD600nm) 1.861 1.830 1.796 1.769 1.731 1.703 1.674 1.633

表6

不同发酵时间多糖的产生与絮凝活性"

检测项目 12 h 16 h 20 h 24 h 28 h 32 h
发酵液pH 5.64 5.62 5.50 5.23 5.51 6.29
黏度(mPa·s) 422 457 503 580 531 479
粗多糖(g·L-1) 17.66 19.92 21.65 23.64 22.92 22.02
生物量(OD600) 1.701 1.697 1.670 1.503 1.576 1.620
FR/% 94.73 96.82 97.90 98.43 97.64 97.20

图1

LF16K多糖生物絮凝剂样品照片"

表7

LF16K多糖生物絮凝剂对黄河水的絮凝数据"

指标 原水样 絮凝处理水样 饮用水指标* 指标 原水样 絮凝处理水样 饮用水指标*
浊度(NTU) 1375 4.62 5 Zn2+(mg/L) 0.515 0.033 1.0
SS(mg/L) 184 0.00 Ni2+(mg/L) 0.019 0.002 0.05
COD(mg/L) 94.67 0.02 3 Hg2+(mg/L) 0.008 0.002 0.001
Fe3+(mg/L) 15.160 0.042 0.3 FR(%) 96.31
Mn2+(mg/L) 4.423 0.043 0.1 pH 7.86 7.82
Cu2+(mg/L) 0.210 0.002 1.0

图2

LF16K多糖生物絮凝剂对黄河水的絮凝照片"

图3

Klebsiella sp.LDX1-1纯化多糖的红外光谱图"

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