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Frontiers of Chemical Science and Engineering

ISSN 2095-0179

ISSN 2095-0187(Online)

CN 11-5981/TQ

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2018 Impact Factor: 2.809

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Front. Chem. Sci. Eng.    2014, Vol. 8 Issue (3) : 330-339    https://doi.org/10.1007/s11705-014-1419-9
RESEARCH ARTICLE
Isolation, identification and primary application of bacteria from putrid alkaline silica sol
Lijie REN,Ye HAN,Shuwen YANG,Xiqian TAN,Jin WANG,Xin ZHAO,Jie FAN,Ting DONG,Zhijiang ZHOU()
School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China
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Abstract

The putrefaction of alkaline silica sol was investigated in this paper. The total colony numbers in three alkaline silica sol samples were 1.47×105, 1.25×104, and 9.45×104 cfu·mL–1, respectively. The salt- and alkali-tolerant strains were isolated and selected using nutrient agar medium at 2.5% salinity and pH 9.5. Basic morphological, physiological and biochemical tests were conducted to confirm the preliminary characterizations of the strains. Based on API 50 CH test and 16S rDNA gene sequence analysis, the isolated strains were finally identified as Exiguobacterium aurantiacum, Cyclobacteriaceae bacterium, Microbacterium sp., Acinetobacter sp., Stenotrophomonas maltophilia and Bacillus thuringiensis. The survivability of the strains under different conditions such as salinities, acidities and temperatures was also studied. Some suitable methods for degerming, such as product pipe steam sterilization and regular canister cleaning, were proposed. To explore the possibility of isolates in industrial application, their alkaline protease and amylase production abilities were preliminarily studied. Five strains produced alkaline protease, whereas two strains produced alkaline amylase. Thus, understanding of the putrefaction on alkaline silica sol would be beneficial for improving industrial production.
Keywords putrid alkaline silica sol      alkaliphile      isolation and identification      sterilization      alkaline protease and alkaline amylase     
Corresponding Author(s): Zhijiang ZHOU   
Issue Date: 11 October 2014
 Cite this article:   
Lijie REN,Ye HAN,Shuwen YANG, et al. Isolation, identification and primary application of bacteria from putrid alkaline silica sol[J]. Front. Chem. Sci. Eng., 2014, 8(3): 330-339.
 URL:  
https://academic.hep.com.cn/fcse/EN/10.1007/s11705-014-1419-9
https://academic.hep.com.cn/fcse/EN/Y2014/V8/I3/330
StrainColony diameterProtuberanceBorderlineSurfaces glossColony textureColony colorCell morphologicalMycoderm
EX0>1mmSwellNeatGlossySmooth and moistLight orangeRod or sphere rod+
EX1<1mmSwellNeatGlossySmooth and moistRedRound or sphere–
EX2<1mmLittle swellNeatGlossySmooth and moistWhiteShort rod or rod+
EX3≈ 1.5mmnoneNeatGlossySoft and moistWhiteShort rod or rod–
EX4<1mmSwellNeatRelusterSoft and dryTransparentRod or sphere rod–
EX5≈ 2mmLittle swellNeatGlossyViscous and moistMilk whiteShort rod or rod++
Tab.1  Morphological characteristics of isolated strains
Test names/sequence numberEX0EX1EX2EX3EX4EX5
Acid productGas productAcid productGas? product?Acid productGas product?Acid productGas? product?Acid productGas ??product???Acid productGas product
Carbohydrates ?fermentationGlucose+–+++++–+– +–
Maltose+++–+++–+– +–
Xylitol–+–+––––––––
Lactose––––––––––––
Galactose–++––––+––––
Sucrose+–+–+–+++– +–
D-fructose+–––+–+–+– +–
Rhamnose––––––––––––
Mannitol––––––––––––
Sorbitol––––––––––––
Glucosamine test+–––+–
Litmus milk testAcid product––––––
Alkaline productpeptonization–+–––+–+––– +
Acid curding+–––– +
Chymosin ?curdingdeoxidation–+–++–+––+– +
Catalase activityUrease activity++–––+–+++– +
Trip sugar-iron ?agar testUpper yellow+––++ +
Lower yellow+–+++ +
Bubble–––+––
Turn black––+++ +
Diffused+–+++ +
Citrate test+–––––
Starch hydrolysis test++–+– +
Tab.2  Physiological and Biochemical characteristics of tested strains
Fig.1  Survivabilities of the six strains at different salinities. The graph’s horizontal axis shows the salinities, and the vertical axis shows the number of bacteria colonies. Each column represents a strain. Vertical bars represent standard deviations of three replicates
Fig.2  Survivabilities of the six strains at different acidities. The graph’s horizontal axis shows different pH values, and the vertical axis shows the number of bacteria colonies. Each column represents a strain. Vertical bars represent standard deviations of three replicates
Fig.3  Survivabilities of the six strains at different temperatures. The graph’s horizontal axis shows different temperatures, and the vertical axis shows the number of bacteria colonies. Each column represents a strain. Vertical bars represent standard deviations of three replicates
NumbersEX0EX1EX2EX3EX4EX5
24 h48 h24 h48 h24 h48 h24 h48 h24 h48 h24 h48 h
0––––––––––––
1V+––V+V+––VV
2––––––––––––
3––––––––––––
4––––––––––––
5–V––V+–V–V–V
6VV––––––––––
7––––––––––––
8––––––––––––
9––––––––––––
10++––––––––––
11++––++++++++
12++––V+–+–V–+
13++––––––––
14––––––––––––
15––––––––––––
16––––––––––––
17––––––––––––
18++––––––––––
19––––––––––––
20––––––––––––
21++––––––––––
22++––++++++++
23++––––––––––
24++––++++++++
25++++++++++++
26++––++++++++
27++––––––––––
28++––++++++++
29––––––––––––
30––––––––––––
31++––++++––++
32++––++++––++
33––––––––––––
34––––––––––––
35––––––––––––
36++–––––––––V
37++––––––––V+
38––––––––––––
39++–––V––––––
40VV––––––––––
41––––––––––––
42––––––––––––
43––––––––––––
44––––––––––––
45––––––––––––
46––––––––––––
47––––––––––––
48––––––––––––
49––––––––––––
Tab.3  Appraisal result of Analytic Products Inc
Fig.4  PCR results of 16S rDNA. The lane of M represents a DGL2000 DNA Marker and the numbers on the left show the length of each band of the marker. The lane of 1 is the negative control. Strains in 2–7 lanes are EX0, EX1, EX2, EX3, EX4, EX5
StrainsSequence length /bpHomology /%Result of identificationGenBank accession number
EX0150797%Exiguobacterium aurantiacumHM030747
EX1138696%Cyclobacteriaceae bacteriumHM140980
EX2139697%Microbacterium sp.FR754552
EX3144098%Acinetobacter sp.HQ704716
EX4151699%Stenotrophomonas maltophiliaFM213389
EX51438100%Bacillus thuringiensisGU391512
Tab.4  Identification of tested strains according to sequence analysis and phenotypic characterization
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