Isolation and Characterization of an Entophytic Ethanol Resistant Bacterium from Sap of <i>Saccharum officinarum</i> for Efficient Fermentation

Rukshika Shalani Hewawasam; Sisira Weliwegamage; Sanath Rajapakse; Subramanium Sotheeswaran

doi:doi:10.11648/j.ajac.20190702.11

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Isolation and Characterization of an Entophytic Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation

Rukshika Shalani Hewawasam, Sisira Weliwegamage, Sanath Rajapakse, Subramanium Sotheeswaran

Published in American Journal of Applied Chemistry (Volume 7, Issue 2)

Received: 23 December 2017 Accepted: 6 March 2018 Published: 9 May 2019

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Abstract

Bio fuel is one of the emerging industries around the world due to arise of crisis in petroleum fuel. Fermentation is a cost effective and eco-friendly process in production of bio-fuel. One major problem in microbial ethanol fermentation is the low resistance of conventional microorganisms to the high ethanol concentrations, which ultimately lead to decrease in the efficiency of the process. In the present investigation, an ethanol resistant bacterium was isolated from sap of Saccharum officinarum (sugar cane). The optimal cultural conditions such as, temperature, incubation period, and microbiological characteristics, morphological characteristics, biochemical characteristics, ethanol tolerance, sugar tolerance were investigated. Isolated microorganism was tolerated to 18% (V/V) of ethanol concentration in the medium and 35% (V/V) glucose concentration in the medium. Biochemical characteristics have revealed as Gram negative, non-motile, negative for Indole test, Methyl Red test, Voges- Proskauer`s test, Citrate Utilization test, and Urease test and positive results for Oxidase test. Sucrose, Glucose, Fructose, Maltose, Dextrose, Arabinose, Raffinose, Lactose, and Sachcharose can be utilized by this particular bacterium. It is a significant feature in effective fermentation. The fermentation process was carried out in glucose medium under optimum conditions; temperature 30°C, and incubated for 72 hours. Maximum ethanol production was recorded as 12.8±0.4% (V/V). Methanol was not detected throughout the fermentation process. This bacterium is especially useful in bio-fuel production due to high ethanol tolerance of this microorganism; it can be used to enhance the fermentation process over conventional microorganisms. Investigations are currently conducted on establishing the identity of the bacterium.

Published in	American Journal of Applied Chemistry (Volume 7, Issue 2)
DOI	10.11648/j.ajac.20190702.11
Page(s)	42-46
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Bacterium, Bio-Fuel, Ethanol Tolerance, Fermentation, Saccharum officinarum

References

[1]	Shapouri H, Duffield JA, Graboski MS. (1995) Estimating the net energy balance of corn ethanol, U. S. Department of Agriculture, Agricultural Economic Report Number 721.
[2]	MacDonald T, Yowell G, McCormack M (2001) US ethanol industry production capacity outlook, California energy commission. Available at: http//www.energy.ca.gov/reports/ 2001-08-29_600-01-017.PDF.
[3]	Gruno M, Vaeljamaee P, Pettersson G, Johansson G (2004) Inhibition of the Trichoderma reesei cellulases by cellobiose is strongly dependent on the nature of the substrate. Biotechnol. Bioengin. 86:503-511.
[4]	Dorothee B, Monteiro ARDS, Costa MMD, Virkajärvi I, Sacon V, Wilhelmsom A (2014) DesinFix TM 135 in fermentation process for bioethanol production. Brazilian Journal of Microbiology 45, 1, 323-325.
[5]	Sasikumar E, Viruthagiri T (2010) Simultaneous Saccharification and Fermentation SSF of Sugarcane Bagasse – Kinetics and Modeling. Int J Chem Biol Engin 32.
[6]	CONAB (2012) Acompanhamento de safra brasileira: cana-deaçúcar, primeiro levantamento, abril/2012. Conab Brasília.
[7]	Backman PA, Sikora RA (2008) Endophytes: An emerging tool for biological control. Biol Cont 46:1-3.
[8]	Thaweenut N, Hachisuka Y, Ando S, Yanagisawa S, Yoneyama T (2011) Two season’ study on nifH gene expression and nitrogen fixation by diazotrophic endophytes in sugarcane (Saccharum spp. hybrids): expression of nifH genes similar to those of rhizobia. Plant Soil 338:435-449.
[9]	Millichip RJ, Doelle HW (1989) Large-scale ethanol production from Milo Sorghum using Zymomonas mobilis. Proc Biochem 24:141-145.
[10]	James EK, Olivares FL (1998) Infection and colonization of sugar cane and other graminaceous plants by endophytic diazotrophs. Critical Reviews in Plant Sciences17, 77–119.
[11]	Sevilla M, Burris RH, Gunapala N, Kennedy C(2001)Comparison of benefit to sugarcane plant growth and 15 N2 incorporation following inoculation of sterile plants with Acetobacter diazotrophicus wild-type and Nif- mutant strains. Molecular Plant Microbe Interactions14:358–366.
[12]	Antwerpen VT, McFarlane SA (2000) The isolation and identification of the causal organism of red stripe symptoms on two sugarcane varieties. 38th Congress of the South African Society for Plant Pathology: 23-28.
[13]	Yoganand S, Luximon AB, Jhurry D, Puchooa D (2012) Isolation of lactic acid bacteria from sugar cane juice and production of lactic acid from selected improved strains, Advances in Bioscience and Biotech3:398-407.
[14]	Tam HM, Diep CN(2014) Isolation, characterization and identification of endophytic bacteria in sugarcane (Saccharum spp. L.) cultivated on soils of the Dong Nai province, Southeast of Vietnam, American Journal of Life Sciences 2(6): 361-368.
[15]	Joseph B, Patra RR, Lawrence R (2007) Characterization of plant growth promoting rhizobacteria associated with chickpea. Int J Plant Prod 1:141-151.
[16]	Osho A (2005) Ethanol and Sugar tolerance of wine yeasts isolated from fermenting cashew apple juice, African Journal of Biotechnology 4 (7):660-662.
[17]	Ekunsanmi TJ, Odunfa SA (1990) Ethanol tolerance, sugar tolerance and invertase activities of some yeasts strains isolated from steep water of fermenting cassava tubers. J. Appl. Bact. 69: 672-675.
[18]	Stackler B, Christensen E N (1974) Quantitative determination of ethanol in wine by Gas Chromatography. Am. J. Enol. Vitic. 25, 202-207.
[19]	Ahmed I, Yokota A, Fujiwara T (2007) A novel highly boron tolerant bacterium, Bacillus boroniphilus sp. nov., isolated from soil, that requires boron for its growth. Extremophiles 11:217-224.
[20]	Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser 41:9-98.
[21]	Manikandan T, Umamaheswari M, Jayakumari M, Maheswari K, Subashree M, Mala P, Sevanthi T., (2010) Bioethanol Production from Cellulosic Materials. Asian J. Sci. Technol. 1:5-11.

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Rukshika Shalani Hewawasam, Sisira Weliwegamage, Sanath Rajapakse, Subramanium Sotheeswaran. (2019). Isolation and Characterization of an Entophytic Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation. American Journal of Applied Chemistry, 7(2), 42-46. https://doi.org/10.11648/j.ajac.20190702.11

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ACS Style

Rukshika Shalani Hewawasam; Sisira Weliwegamage; Sanath Rajapakse; Subramanium Sotheeswaran. Isolation and Characterization of an Entophytic Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation. Am. J. Appl. Chem. 2019, 7(2), 42-46. doi: 10.11648/j.ajac.20190702.11

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AMA Style

Rukshika Shalani Hewawasam, Sisira Weliwegamage, Sanath Rajapakse, Subramanium Sotheeswaran. Isolation and Characterization of an Entophytic Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation. Am J Appl Chem. 2019;7(2):42-46. doi: 10.11648/j.ajac.20190702.11

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@article{10.11648/j.ajac.20190702.11,
  author = {Rukshika Shalani Hewawasam and Sisira Weliwegamage and Sanath Rajapakse and Subramanium Sotheeswaran},
  title = {Isolation and Characterization of an Entophytic Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation},
  journal = {American Journal of Applied Chemistry},
  volume = {7},
  number = {2},
  pages = {42-46},
  doi = {10.11648/j.ajac.20190702.11},
  url = {https://doi.org/10.11648/j.ajac.20190702.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajac.20190702.11},
  abstract = {Bio fuel is one of the emerging industries around the world due to arise of crisis in petroleum fuel. Fermentation is a cost effective and eco-friendly process in production of bio-fuel. One major problem in microbial ethanol fermentation is the low resistance of conventional microorganisms to the high ethanol concentrations, which ultimately lead to decrease in the efficiency of the process. In the present investigation, an ethanol resistant bacterium was isolated from sap of Saccharum officinarum (sugar cane). The optimal cultural conditions such as, temperature, incubation period, and microbiological characteristics, morphological characteristics, biochemical characteristics, ethanol tolerance, sugar tolerance were investigated. Isolated microorganism was tolerated to 18% (V/V) of ethanol concentration in the medium and 35% (V/V) glucose concentration in the medium. Biochemical characteristics have revealed as Gram negative, non-motile, negative for Indole test, Methyl Red test, Voges- Proskauer`s test, Citrate Utilization test, and Urease test and positive results for Oxidase test. Sucrose, Glucose, Fructose, Maltose, Dextrose, Arabinose, Raffinose, Lactose, and Sachcharose can be utilized by this particular bacterium. It is a significant feature in effective fermentation. The fermentation process was carried out in glucose medium under optimum conditions; temperature 30°C, and incubated for 72 hours. Maximum ethanol production was recorded as 12.8±0.4% (V/V). Methanol was not detected throughout the fermentation process. This bacterium is especially useful in bio-fuel production due to high ethanol tolerance of this microorganism; it can be used to enhance the fermentation process over conventional microorganisms. Investigations are currently conducted on establishing the identity of the bacterium.},
 year = {2019}
}

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TY - JOUR
T1 - Isolation and Characterization of an Entophytic Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation
AU - Rukshika Shalani Hewawasam
AU - Sisira Weliwegamage
AU - Sanath Rajapakse
AU - Subramanium Sotheeswaran
Y1 - 2019/05/09
PY - 2019
N1 - https://doi.org/10.11648/j.ajac.20190702.11
DO - 10.11648/j.ajac.20190702.11
T2 - American Journal of Applied Chemistry
JF - American Journal of Applied Chemistry
JO - American Journal of Applied Chemistry
SP - 42
EP - 46
PB - Science Publishing Group
SN - 2330-8745
UR - https://doi.org/10.11648/j.ajac.20190702.11
AB - Bio fuel is one of the emerging industries around the world due to arise of crisis in petroleum fuel. Fermentation is a cost effective and eco-friendly process in production of bio-fuel. One major problem in microbial ethanol fermentation is the low resistance of conventional microorganisms to the high ethanol concentrations, which ultimately lead to decrease in the efficiency of the process. In the present investigation, an ethanol resistant bacterium was isolated from sap of Saccharum officinarum (sugar cane). The optimal cultural conditions such as, temperature, incubation period, and microbiological characteristics, morphological characteristics, biochemical characteristics, ethanol tolerance, sugar tolerance were investigated. Isolated microorganism was tolerated to 18% (V/V) of ethanol concentration in the medium and 35% (V/V) glucose concentration in the medium. Biochemical characteristics have revealed as Gram negative, non-motile, negative for Indole test, Methyl Red test, Voges- Proskauer`s test, Citrate Utilization test, and Urease test and positive results for Oxidase test. Sucrose, Glucose, Fructose, Maltose, Dextrose, Arabinose, Raffinose, Lactose, and Sachcharose can be utilized by this particular bacterium. It is a significant feature in effective fermentation. The fermentation process was carried out in glucose medium under optimum conditions; temperature 30°C, and incubated for 72 hours. Maximum ethanol production was recorded as 12.8±0.4% (V/V). Methanol was not detected throughout the fermentation process. This bacterium is especially useful in bio-fuel production due to high ethanol tolerance of this microorganism; it can be used to enhance the fermentation process over conventional microorganisms. Investigations are currently conducted on establishing the identity of the bacterium.
VL - 7
IS - 2
ER -

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Author Information

Rukshika Shalani Hewawasam

Post Graduate Institute of Science, University of Peradeniya, Peradeniya, Sri Lanka
Sisira Weliwegamage

College of Chemical Sciences, Institute of Chemistry Ceylon, Rajagiriya, Sri Lanka
Sanath Rajapakse

Department of Molecular Biology and Biotechnology, Faculty of Science, University of Peradeniya, Sri Lanka
Subramanium Sotheeswaran

College of Chemical Sciences, Institute of Chemistry Ceylon, Rajagiriya, Sri Lanka

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Rukshika Shalani Hewawasam, Sisira Weliwegamage, Sanath Rajapakse, Subramanium Sotheeswaran. (2019). Isolation and Characterization of an Entophytic Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation. American Journal of Applied Chemistry, 7(2), 42-46. https://doi.org/10.11648/j.ajac.20190702.11

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ACS Style

Rukshika Shalani Hewawasam; Sisira Weliwegamage; Sanath Rajapakse; Subramanium Sotheeswaran. Isolation and Characterization of an Entophytic Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation. Am. J. Appl. Chem. 2019, 7(2), 42-46. doi: 10.11648/j.ajac.20190702.11

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AMA Style

Rukshika Shalani Hewawasam, Sisira Weliwegamage, Sanath Rajapakse, Subramanium Sotheeswaran. Isolation and Characterization of an Entophytic Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation. Am J Appl Chem. 2019;7(2):42-46. doi: 10.11648/j.ajac.20190702.11

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@article{10.11648/j.ajac.20190702.11,
  author = {Rukshika Shalani Hewawasam and Sisira Weliwegamage and Sanath Rajapakse and Subramanium Sotheeswaran},
  title = {Isolation and Characterization of an Entophytic Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation},
  journal = {American Journal of Applied Chemistry},
  volume = {7},
  number = {2},
  pages = {42-46},
  doi = {10.11648/j.ajac.20190702.11},
  url = {https://doi.org/10.11648/j.ajac.20190702.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajac.20190702.11},
  abstract = {Bio fuel is one of the emerging industries around the world due to arise of crisis in petroleum fuel. Fermentation is a cost effective and eco-friendly process in production of bio-fuel. One major problem in microbial ethanol fermentation is the low resistance of conventional microorganisms to the high ethanol concentrations, which ultimately lead to decrease in the efficiency of the process. In the present investigation, an ethanol resistant bacterium was isolated from sap of Saccharum officinarum (sugar cane). The optimal cultural conditions such as, temperature, incubation period, and microbiological characteristics, morphological characteristics, biochemical characteristics, ethanol tolerance, sugar tolerance were investigated. Isolated microorganism was tolerated to 18% (V/V) of ethanol concentration in the medium and 35% (V/V) glucose concentration in the medium. Biochemical characteristics have revealed as Gram negative, non-motile, negative for Indole test, Methyl Red test, Voges- Proskauer`s test, Citrate Utilization test, and Urease test and positive results for Oxidase test. Sucrose, Glucose, Fructose, Maltose, Dextrose, Arabinose, Raffinose, Lactose, and Sachcharose can be utilized by this particular bacterium. It is a significant feature in effective fermentation. The fermentation process was carried out in glucose medium under optimum conditions; temperature 30°C, and incubated for 72 hours. Maximum ethanol production was recorded as 12.8±0.4% (V/V). Methanol was not detected throughout the fermentation process. This bacterium is especially useful in bio-fuel production due to high ethanol tolerance of this microorganism; it can be used to enhance the fermentation process over conventional microorganisms. Investigations are currently conducted on establishing the identity of the bacterium.},
 year = {2019}
}

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