Article Sidebar

Submitted
Feb 22, 2021
Accepted
Mar 18, 2021
Published
Mar 27, 2021
Download
pdf
Statistic
Read Counter : 409 Download : 268

Downloads

Download data is not yet available.

Main Article Content

Abstract

Cellulolytic microbes produce cellulase enzymes that can degrade cellulose. Cellulolytic microbes are found in many habitats, including in cattle waste (bio-slurry), which is result of hydrolysis of cattle feed ingredients sourced from cellulose. Cellulase enzymes play a role in the process of hydrolyzing cellulose into glucose. This study aims to isolate cellulolytic microbes from bio-slurry and measure activity of cellulase enzymes in cellulolytic microbes isolated from bio-slurry. We are taking bio-slurry samples with 5 sample points and isolating the microbes in Carboxyl Methyl Cellulose (CMC) medium, cellulase enzyme activity qualitative test with Congo Red Assay, and perform quantitative test with the DNS Assay. Microbial isolates which shows positive results for cellulolytic activity in the qualitative test called SIIC1, SIIIC5, SIIC3. In the quantitative test can be seen based on Optical Density (OD) for cell lysate, SIIC1 0.298; SIIC3 0.186; SIIIC5 0.247; and a 0.332 for blank solution. Whereas supernatant obtain SIIC1 value of 0.237; SIIC3 0.212; SIIIC5 0.198; and 0.195 for blank solution. Based on the results, it can be concluded that the cellulolytic activity of microbes in bio-slurry based on OD values shows good results in supernatant, but it’s not significant enough in the cell lysate.

Keywords

Cellulolytic microbe cellulase enzyme bio-slurry Carboxyl Methyl Cellulose (CMC)

Article Details

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

How to Cite
1.
Muammar A, Manullang M, Arjuna M, Retnaningrum E. Isolation of Cellulolytic Microbes from Bio-Slurry. EKSAKTA [Internet]. 2021Mar.27 [cited 2024Apr.25];22(1):27-34. Available from: https://eksakta.ppj.unp.ac.id/index.php/eksakta/article/view/257

References

  1. Bernalier A, Fonty G, Bonnemoy F. Degradation and fermentation of cellulose by the rumen anaerobic fungi in axenic cultures or in association with cellulolytic bacteria. Current Microbiology. 1992. 25: 143–148.
  2. Lehninger. Dasar Dasar Biokimia. Jakarta: PT. Gramedia Pustaka Utama; 1992.
  3. Didya S. Kemelimpahan Bakteri Selulolitik di Muara Sungai Anyar Surabaya dan Bancaran Bangkalan. Skripsi Fakultas Peri-kanan dan Kelautan Universitas Airlangga. Surabaya. 2014. P. 18.
  4. Hartanti Y and Putri C. Pedoman Pengguna dan Pengawas: Pengelolaan dan Pemanfaatan Bio-slurry. Jakarta: PT. Gramedia Pustaka; 2013.
  5. Saha BC. Lignocellulose Biodegradation and Application in Biotechnology. US Government Work. American Chemical Society. 2004. pp.2-14.
  6. Ruzki AA. Bio-Degradasi Selulosa Hasil Bio-Pretreatment Jerami Padi Secara Fermen-tasi Padat Menggunakan Isolat Actinomycetes Acp-1 dan Acp-7. Fakultas MIPA Universitas Lampung. 2013. P.6.
  7. Han Q, Rossow W, Welch R, White A and Chou J. Characterization of AGT1 encoding a general alpha-glucoside transporter from Saccharomyces. Mol Microbiol. 1995. 17(6): 107-193.
  8. Ibrahim A and El-diwany. Isolation and Identification of New Cellulases Producing Thermophilic Microbe from an Egyptian Hot Spring and Some Properties of the Crude Ezyme. Australian Journal of Basic and Applied Sciences. 2007. 1(4): 473-478.
  9. Thapa S, Mishra J, Arora N. Microbial cellulolytic enzymes: diversity and biotechnology with reference to lignocellulosic biomass degradation. Rev Environ Sci Biotechnol. 2020. 19:621–648.
  10. Ramadhan ML. Analisis Potensi dan Karak-terisasi Molukuler Gen 16S rRNA Bakteri Selulolitik yang Diisolasi dari Mikroalga Euceuma sp. dan Sargassum sp. sebagai Penghasil Selulase. Jurnal Perikanan dan Kelautan UNPAD Bandung. 2012. 3(3): 61-67.
  11. Weiping Yang, Fanxu Meng, Jiayin Peng, Peng Han, Fang Fang, Li Ma, Binyun Cao. Isolation and identification of a cellulolytic bacterium from the Tibetan pig's intestine and investigation of its cellulase production. Electronic Journal of Biotechnology. 2014. 17:6262-267.
  12. Tortora GJ. Microbiology: An Introduction. New York: The Benjamin. 2010.
  13. Fardiaz S. Mikrobiologi Pangan I. Jakarta: Gramedia Pustaka Utama; 1992.
  14. Waluyo L. Teknik Metode Dasar Dalam Mikrobiologi. Malang: Universitas Muhammadiyah
  15. Malang Press; 2008.
  16. Gohel HR, Contractor CN, Ghosh SK and Braganza VJ. A comparative study of various staining techniques for determination of extra cellular cellulase activity on Carboxy Methyl Cellulose (CMC) agar plates. International Journal of Current Microbiology and Aplied Sciences. 2014. 3:261–266.
  17. Narendra Naik Deshavath, Gourab Mukherjee, Vaibhav V Goud, Venkata Dasu Veeranki, Chivukula V Sastri. Pitfalls in the 3, 5-dinitrosalicylic acid (DNS) assay for the reducing sugars: Interference of furfural and 5-hydroxymethylfurfural. International Journal of Biological Macromolecules. 2020. 156: 180-185.
  18. Pelczar, Chan and Krieg. Microbiology. New York: Mc-Graw Hill Book Company; 1999.
  19. Carder JH. Detection and quantitation of cellulase by Congo red staining of substrates in a cup-plate diffusion assay. Anal Biochem. 1986.153 (1):75-9.
  20. Alexander VG, Elena GK, Arkady PS. Comparison of Two Methods for Assaying Reducing Sugars in the Determination of Carbohydrase Activities. International Journal of Analytical Chemistry. 2011.
  21. Kasana RC, Salwan R, Dhar H. A Rapid and Easy Method for the Detection of Microbial Cellulases on Agar Plates Using Gram’s Iodine. Curr Microbiol. 2008. 57:503–507.
  22. Muammar A, Fajrin SAR, Retnaningrum E. Cellobiohydrolase A (CBHA) gene cloning from Aspergillus niger to the yeast expression vector as a stages to create cellulosic ethanol strain. AIP Conference Proceedings 2260, 060003. 2020. https://doi.org/10.1063/5.0016145
  23. Anand, Vennison, Sankar, Prabhu, Vasan and Vendan. Isolation and Characterization of Bacteria from the Gut of Bombyx Mori that Degrade Cellulose, Xylan, Pectin and Starch and Their Impact on Digestion. Journal of Insect Science. 2009. 10(107):1-20.
  24. Zverlova VV. Hol W and Schwarz H. Enzymes for Digestion of Cellulose and other Polysaccharides in the Gut of Longhorn Beetle Larvae, Rhagium inquisitor L. (Col., Cerambycidae). International Biodeterioration & Biodegradation. 2003. 51(2):175-179.
  25. Lynd LR, Weimer WH and Pretorius IS. Microbial Cellulose Utilization: Fundamentals and Biotechnology. Microbiol Mol.Biol. 2002. 66(3):506-577.
  26. Zhang J. Characterization of the transport mechanism and permeant binding profile of the uridine permease Fui1p of Saccharomyces cerevisiae. Biology Chemistry Journal. 2006. 8(38): 210-2.
  27. Fikrinda N. Isolasi dan Karakterisasi Bakteri Penghasil Selulase Ekstermofilik dari Ekosistem Air Hitam. IPB. Bogor. 2000.
  28. Nurmayani D. Isolasi dan Uji Potensi Mikrooganisme Selulolitik Asal Tanah Gambut dan Kayu Sedang Melapuk Dalam Mendekomposisikan Kayu. Universitas Sumatera Utara. 2007.