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

Previous Articles    

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

RichHTML

12

PDF (PC)

852

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"

1 ISMAIL B , NAMPOOTHIRI K M . Production, purification and structural characterization of an exopolysaccharide produced by a probiotic Lactobacillus plantarum MTCC 9510[J]. Archives of Microbiology, 2010, 192 (12): 1049- 1057.
doi: 10.1007/s00203-010-0636-y
2 DE VUYST L , DEGEEST B . Heteropolysaccharides from lactic acid bacteria[J]. FEMS Microbiology Reviews, 1999, 23 (2): 153- 177.
doi: 10.1016/S0168-6445(98)00042-4
3 LAWS A , GU Yucheng , MARSHALL V . Biosynthesis, characterization, and design of bacterial exopolysaccharides from lactic acid bacteria[J]. Biotechnology Advances, 2001, 19 (8): 597- 625.
doi: 10.1016/S0734-9750(01)00084-2
4 ARENA A , MAUGERI T L , PAVONE B. , et al. Antiviral and immunoregulatory effect of a novel exopolysaccharide from a marine thermotolerant Bacillus licheniformis[J]. International Immunopharmacology, 2006, 6 (1): 8- 13.
5 SARAVANAN C , KAVITAKE D , KANDASAMY S , et al. Production, partial characterization and antioxidant properties of exopolysaccharide a-D-glucan produced by Leuconostoc lactis KC117496 isolated from an idli batter[J]. Journal of Food Science and Technology, 2018.
doi: 10.1007/s13197-018-3469-3
6 ABINAYAL M , VASEEHARAN B , DIVYAL M , et al. Structural characterization of Bacillus licheniformis Dahb1 exopolysaccharide—antimicrobial potential and larvicidal activity on malaria and Zika virus mosquito vectors[J]. Environmental Science and Pollution Research, 2018.
doi: 10.1007/s11356-018-2002-6
7 WANG Yanping , AHMED Z , WU Feng , et al. Physicochemical properties of exopolysaccharide produced by Lactobacillus kefiranofaciens ZW3 isolated from Tibet kefir[J]. International Journal of Biological Macromolecules, 2008, 43 (3): 283- 288.
doi: 10.1016/j.ijbiomac.2008.06.011
8 PISHCHIK V N , CHERNYAEVA I I , SEMENOVA E A , et al. Metabolic features of bacteria of the Genus Klebsiella[J]. Microbiology, 1997, 66 (1): 54- 59.
9 BRYAN B A , LINHARDT R J , DANIELS L . Variation in composition and yield of exopolysaccharides produced by Klebsiella sp. strain K32 and Acinetobacter calcoaceticus BD[J]. Applied and Environmental Microbiology, 1986, 51 (6): 1304- 1308.
doi: 10.1128/AEM.51.6.1304-1308.1986
10 PEIRIS P S , DLAMINI A M , BAVOR H J . Optimization of bioprocess conditions for exopolysaccharide production by Klebsiella oxytoca[J]. World Journal of Microbiology & Biotechnology, 1998, 14: 917- 919.
11 NAKATA K , KURANE R . Production of an extracellular polysaccharide bioflocculant by Klebsiella pneumoniae[J]. Bioscience Biotechnology and Biochemistry, 1999, 63 (12): 2064- 2068.
doi: 10.1271/bbb.63.2064
12 YIN Yajie , TIAN Zuning , TANG Wei , et al. Production and characterization of high efficiency bioflocculant isolated from Klebsiella sp. ZZ-3[J]. Bioresource Technology, 2014, 17: 336- 342.
13 DUBOIS M , GILLES K A , HAMILTON J K , et al. Colorimetric method for determination of sugars and related substances[J]. Analytical Chemistry, 1956, 28 (3): 350- 356.
14 栾兴社. 水处理多糖生物絮凝剂研究[M]. 北京: 科学出版社, 2019: 165- 166.
LUAN Xingshe . Study on biological flocculant of polysaccharide in water treatment[M]. Beijing: Science Press, 2019: 165- 166.
15 栾兴社, 王桂宏, 于伟正, 等. 微生物絮凝剂产生菌节杆菌LF-Tou2的培养和絮凝条件研究[J]. 现代化工, 2004, 24 (6): 43- 45.
LUAN Xingshe , WANG Guihong , YU Weizheng , et al. Study on culture and flocculation conditions of Arthrobacter sp. LF-Tou2 which can produce microbial flocculant[[J]. Modern Chemical Industry, 2004, 24 (6): 43- 45.
16 栾兴社.节杆菌LF-Tou2的分离和产生生物絮凝剂的特征研究[D].济南:山东大学, 2005.
LUAN Xingshe. Isolation of Arthrobacter LF-Tou2 and study on the characteristics of bioflocculant production[D]. Jinan: Shandong University, 2005.
17 中国国家环保总局. 水和废水检测分析方法[M]. 4版 北京: 中国环境科学出版社, 2002.
State Environmental Protection Administration of China . Water and wastewater detection and analysis methods[M]. 4th Edition Beijing: China Environmental Science Press, 2002.
18 罗士津, 瞿明仁. 松针粉的研究及应用进展[J]. 饲料工业, 2007, 28 (3): 54- 57.
LUO Shijin , QU Mingren . Research and application progress of pine needle powder[J]. Feed Industry, 2007, 28 (3): 54- 57.
19 RASULOV B A , LI Li , LIU Yonghong , et al. Production, characterization and structural modification of exopolysaccharide-based bioflocculant by Rhizobium radiobacter SZ4S7S14 and media optimization[J]. Biotechnology, 2017, 7 (3): 179- 188.
20 LUAN Xingshe. Klebsiella and method for preparing microbial flocculant with same: US 10, 150, 943, B2[P].2018-12-11.
[1] ZHANG Jie, ZHANG Wei-Ming, SHI Jin-Song, SUN Da-Feng, ZHU Chang-Ling. Rheological properties of Blettila striata polysaccharide solutions [J]. J4, 2010, 45(3): 15-18.
[2] . Effect of intraperitoneal injection of indigowoad ro ot polysaccharide on
antioxidant enzyme activity and MDA content in the liver of Pelodiscus sinensis [J]. J4, 2009, 44(7): 5-7.
[3] . Modeling in fedbatch fermentation and its parameter 
identification based on HPSO algorithm [J]. J4, 2009, 44(7): 71-78.
[4] . Pnitroaniline biosorption by a novel exopolysaccharide from the deepsea mesophilic bacterium Wangia profunda (SMA87) [J]. J4, 2009, 44(5): 33-39.
[5]

WU Rong-rong,WANG Li-ping,GAO Li-li,LIU Bai-qu,ZHANG Bo-lin *

.

Exopolysaccharide production and lyophilization preparation from Streptococcus thermophilus ST

[J]. J4, 2008, 43(7): 88-96 .
[6] LI Ai-li,HUO Gui-cheng*,DENG Kai-bo .

Effect of yeast extract concentration on the batch fermentation kinetics of
Lactobacillus delbrueckii subsp.bulgaricus KLDS 1.9201

[J]. J4, 2008, 43(7): 23-27 .
[7] CHEN Jun-liang,YANG Li-jie,HUO Gui-cheng* .

Identification of α-galactosidase-producing lactic acid bacteria and their fermentation performance

[J]. J4, 2008, 43(7): 28-32 .
[8] SHI Lei,FU You-li and CHEN Kao-shan* . Separation, purification and component research of a polysaccharide from the roots of Cudrania tricuspidata (Carr.) Bur. [J]. J4, 2007, 42(5): 74-78 .
[9] CHEN Yu-juan,JI Ai-guo,* and GAO Pei-ji . Purification and characterization of a new kind of antimicrobial peptide by fungi-penicillium sp.M03 [J]. J4, 2007, 42(5): 64-68 .
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] MENG Xiang-bo1, ZHANG Li-dong1, DU Zi-ping2. Investment and reinsurance strategy for insurers under #br# mean-variance criterion with jumps#br#[J]. JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE), 2014, 49(05): 36 -40 .
[2] PENG Zhen-hua, XU Yi-hong*, TU Xiang-qiu. Optimality conditions for weakly efficient elements of nearly preinvex set-valued optimizaton#br#[J]. JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE), 2014, 49(05): 41 -44 .
[3] HU Ming-Di, SHE Yan-Hong, WANG Min. Topological properties of  three-valued   logic  metric space[J]. J4, 2010, 45(6): 86 -90 .
[4] LIU Zhan-jie1, MA Ru-ning1, ZOU Guo-ping1, ZHONG Bao-jiang2, DING Jun-di 3. An algorithm for color image segmentation based on region growth[J]. J4, 2010, 45(7): 76 -80 .
[5] YUAN Hun-ping . Schur factorization and normal matrices factorization of row (column) symmetric matrices[J]. J4, 2007, 42(10): 123 -126 .
[6] LIU Feng-qin,XIA Hai-rui,RAN Dong-gang,ZHAO Peng,SAN Shang-qian,LING Zong-cheng,GAO Wen-lan, . The Raman spectra analysis of BaWO4 crystal[J]. J4, 2006, 41(5): 115 -118 .
[7] HE Hai-lun， CHEN Xiu-lan* . Circular dichroism detection of the effects of denaturants and buffers on the conformation of cold-adapted protease MCP-01 and  mesophilic protease BP01[J]. JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE), 2013, 48(1): 23 -29 .
[8] WANG Bi-yu, CAO Xiao-hong*. The perturbation for the Browder’s theorem of operator matrix#br#[J]. JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE), 2014, 49(03): 90 -95 .
[9] DING Chao1, 2, YUAN Chang-an1, 3, QIN Xiao1, 3. A prediction algorithm for multi-data streams  based on GEP[J]. J4, 2010, 45(7): 50 -54 .
[10] GUO Qiao-jin, DING Yi, LI Ning. A context based method for ROI detection in digitized mammograms[J]. J4, 2010, 45(7): 70 -75 .
Copyright 2018 © Editorial office of JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE)
Supported by Beijing Magtech Co.Ltd