An Extracellular Pectinase from ISH16 Bacteria Isolated Induced by Coffee Pulp Waste Substrate

Kahar Muzakhar(1*), Farah Salma Elida(2), Ramdhan Putrasetya(3), Siswoyo Siswoyo(4), Rudju Winarsa(5), Hidayat Teguh Wiyono(6)

(1) Biology Department, Faculty of Mathematic and Natural Sciences, Universitas Jember, Jl. Kalimantan 37, Jember, East Java, Indonesia, 68121, Indonesia
(2) Biology Department, Faculty of Mathematic and Natural Sciences, Universitas Jember, Jl. Kalimantan 37, Jember, East Java, Indonesia, 68121, Indonesia
(3) Biology Department, Faculty of Mathematic and Natural Sciences, Universitas Jember, Jl. Kalimantan 37, Jember, East Java, Indonesia, 68121, Indonesia
(4) Chemistry Department, Faculty of Mathematic and Natural Sciences, Universitas Jember, Jl. Kalimantan 37, Jember, East Java, 68121, Indonesia
(5) Biology Department, Faculty of Mathematic and Natural Sciences, Universitas Jember, Jl. Kalimantan 37, Jember, East Java, Indonesia, 68121, Indonesia
(6) Biology Department, Faculty of Mathematic and Natural Sciences, Universitas Jember, Jl. Kalimantan 37, Jember, East Java, Indonesia, 68121,  
(*) Corresponding Author


An α-1,4-glycosidic bonds galactoses pectin, mainly composed of a D-galacturonic acid chain, are important biomaterial widely used in industries. Utilizing this material, a bioprocess, including the biocatalysis pectinase, is often needed. Pectinase production was optimized in 7 days SSF at 37°C, and the pectinase activities were daily measured by the method of Somogy-Nelson. The optimum pectinase production was 0.166 U/ml on the fourth day SSF. Purification using open column ion exchange chromatography DEAE cellulose DE-52 resulted in 1030.9 folds of pectinase purity with a yield of 25.9%. The enzyme was at optimal activity at pH six and attended stable in the pH range of 5.5-8, while optimal activity at a temperature of 50°C and was stable in the range of 30-45°C. The pectinase activity increased by 120% with the addition of 10 mM Mg2+, and 95% retained when 10 mM Ca2+ was added. The presence of 10 mM Na+, K+, and Fe2+ resulted in a slight effect of activity at 85%, 83%, and 78%. However, it was strongly inhibited by 10 mM Al3+ and retained 25%. Based on the results above, the microbial utilization of coffee pulp waste by ISH16 bacteria pectinolytic is one opportunity to produce valuable pectinase with low-cost production, so comprehensive examination in large-scale production is needed too. In this paper, all research detail steps were described.


coffee pulp, ISH16 bacteria isolate, pectinase, purification, solid state fermentation

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