JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE) ›› 2020, Vol. 55 ›› Issue (7): 103-110.doi: 10.6040/j.issn.1671-9352.0.2019.667

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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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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

CLC Number: 

  • Q93

Table 1

Effect of carbon source on polysaccharide production"

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

Table 2

Effect of mass fraction of isomaltooligosaccharide in mixed sugar on polysaccharide production"

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

Table 3

Effect of nitrogen source on polysaccharide production"

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

Table 4

Effects of growth compound factors on the production of polysaccharides"

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

Table 5

Effect of Triton X-100 on polysaccharide production"

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

Table 6

Production and flocculation activity of polysaccharide at different fermentation time"

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

Fig.1

Photo of sample of LF16K polysaccharide bioflocculant"

Table 7

Flocculation data of LF16K polysaccharide bioflocculant on the Yellow River water"

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

Fig.2

Flocculation photo of LF16K polysaccharide bioflocculant on Yellow River water"

Fig.3

IR spectra of purified polysaccharide from Klebsiella sp. LDX1-1"

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