Biotechnology and Bioprocess Engineering 2020; 25(1): 132-140  
Enhancement of Hydrolysis and Biogas Production of Primary Sludge by Use of Mixtures of Protease and Lipase
Jovale Vincent Tongco1, Sangmin Kim1, Baek-Rock Oh2, Sun-Yeon Heo2, Joonyeob Lee1, and Seokhwan Hwang1,*
1Division of Environmental Science and Engineering, Pohang University of Science and Technology (POSTECH), Pohang 37673, Korea
2Microbial Biotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeong-eup 56212, Korea
Correspondence to: Seokhwan Hwang*
Division of Environmental Science and Engineering, Pohang University of Science and Technology (POSTECH), Pohang 37673, Korea
Tel: +82-54-279-2282; Fax: +82-54-279-8299
Received: July 24, 2019; Revised: September 9, 2019; Accepted: September 21, 2019; Published online: February 29, 2020.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
This study aims to improve the hydrolysis and degradation of primary sludge by using wild-type enzymes (protease and lipase) and establishing the optimal enzymatic cocktail ratio. Primary sludge from three wastewater treatment plants (WWTPs) in Korea (Ulsan, Pohang, and Busan) were subjected to enzymatic hydrolysis. Protease and lipase were isolated from enzyme-producing microorganisms cultured from secondary sludge collected at 8 different digester sites in Korea. Primary sludge degradation through enzymatic hydrolysis was monitored by measuring the reduction in the volatile suspended solids (VSS) content of the sludge and enzyme cocktail mix for 72 h at 40oC and pH 7.0. The enzymatic cocktail of Ulsan primary sludge treated with protease to lipase at a ratio of 1:3 was found to be optimal at 33.3% VSS reduction. Biochemical methane potential (BMP) tests were employed to the optimal enzyme cocktail to measure the potential of the hydrolyzed substrate for further degradation (VSS reduction) and bioconversion to biogas using 125 mL serum bottles as anaerobic reactors for 30 days. BMP tests showed that there was an increase in biogas production by 84.1%, methane production by 89.8%, and methane yield by 9.6%. Methane production rate was also increased. The significant VSS concentration reduction and higher biogas and methane yield of the enzyme-treated primary sludge correlate to the fact that the complex polymeric organic materials were degraded leading to efficient utilization by the microorganisms in the anaerobic digestion process.
Keywords: primary sludge, enzyme cocktail, hydrolysis, anaerobic digestion, protease, lipase

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