Proceedings of the Zoological Society

, Volume 72, Issue 2, pp 160–170 | Cite as

New Extracellular Polymeric Substance Producing Enteric Bacterium from Earthworm, Metaphire posthuma: Modulation Through Culture Conditions

  • Jayanta Kumar BiswasEmail author
  • Anurupa Banerjee
  • Santanu Majumder
  • Nanthi Bolan
  • Balaji Seshadri
  • Madhab Chandra Dash
Research Article


An extracellular polymeric substance (EPS) producing microorganism, identified and characterized as Bacillus licheniformis strain KX657843 based on 16S rRNA sequencing and phylogenetic analysis, was isolated from earthworms (Metaphire posthuma). The bacteria was found to be uniquely influenced by the amino acid l-asparagine monohydrate. The organism produced the maximum yield of 2.7 g L−1 EPS in the presence of sucrose supplemented with l-asparagine monohydrate while no EPS was produced in absence of the amino acid. The order of growth and EPS production in presence of different carbon substrates supplemented with l-asparagine monohydrate was observed as sucrose > glucose > glycerol > mannitol > citrate > cellulose > starch. l-asparagine monohydrate could serve as the sole nitrogen source for the organism but it alone cannot satisfy the requisite carbon demand for growth and EPS production. EPS production showed a positive correlation with sucrose concentration but a negative correlation with citrate levels. Analysis of extracted EPS using Fourier transform infrared spectroscopy and measurement of its zeta potential revealed the chemical composition and anionic nature of the EPS. The bacterium produced 119.235 IU mL−1 extracellular l-asparaginase. This is the first report of an l-asparagine monohydrate dependent EPS producing Bacillus licheniformis from the gut of the earthworm, Metaphire posthuma.


Extracellular polymeric substances Earthworm Gut bacteria l-Asparagine monohydrate Carbon substrate 



The study was financially supported by Department of Science and Technology (DST), Government of India in the form of providing INSPIRE Fellowship (Code No.: 1F131101) to Anurupa Banerjee for carrying out research work under the supervision of Dr. Jayanta Kumar Biswas. The Authors duly acknowledge the infrastructural support provided by Department of Ecological Studies, University of Kalyani as well as the facility available under the DST-PURSE programme. SM is thankful to the Fulbright-Nehru Post-Doctoral Fellowship Programme (Award No. 2203/FNPDR/2016) from the United States-India Educational Foundation (USIEF). Professor Mary Beth Kirkham, Department of Agronomy, Kansas State University Manhattan, USA made constructive comments on the manuscript and rendered help with its English composition. Authors express their gratitude to Dr. Amit Chowdhury, an earthworm expert for his help in identifying the earthworm species. Authors are also thankful to Mr. Prabuddha Sur, Department of Chemistry, University of Kalyani for helping in the FTIR analysis.

Compliance with ethical standards

Conflict of interest

The authors have no financial/commercial conflicts of interest.


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Copyright information

© Zoological Society, Kolkata, India 2017

Authors and Affiliations

  • Jayanta Kumar Biswas
    • 1
    • 2
    Email author
  • Anurupa Banerjee
    • 1
  • Santanu Majumder
    • 1
    • 3
  • Nanthi Bolan
    • 4
    • 5
  • Balaji Seshadri
    • 4
    • 5
  • Madhab Chandra Dash
    • 6
  1. 1.Department of Ecological StudiesUniversity of KalyaniKalyani, NadiaIndia
  2. 2.International Centre for Ecological EngineeringUniversity of KalyaniKalyaniIndia
  3. 3.Department of Geology and GeophysicsTexas A&M UniversityCollege StationUSA
  4. 4.Global Centre for Environmental Remediation (GCER)University of NewcastleNewcastleAustralia
  5. 5.Cooperative Research Centre for Contamination Assessment and Remediation of the Environment (CRC CARE)CallaghanAustralia
  6. 6.Sambalpur UniversitySambalpurIndia

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