Isolation, Screening, Production and Characterization of Xylanase from Air Panas Hot Spring Bacteria (Soil)

Lim, Jian Wen (2016) Isolation, Screening, Production and Characterization of Xylanase from Air Panas Hot Spring Bacteria (Soil). Final Year Project (Bachelor), Tunku Abdul Rahman University College.

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Abstract

The study of Xylanase becomes one of the most important studies due to its large potential of application in paper pulp, food industry and also the conversion of agro wastes. The aim of the present study was to isolate and characterize xylanase producing bacteria from Air Panas Hot Springs, Kuala Lumpur, which was capable of producing xylanase at high temperature. Different types of samples were obtained such as water, biomass and soil from the hot spring. Generally, soil samples had provided higher isolation rates compared to water and biomass samples. Xylanase-producing bacteria were isolated from soil sample. SM03 Isolate with the largest clear zone up to 1.0cm were selected and subjected to bacteria identification and xylanase production. SM03 Isolate was grown in shaking flasks with oat spelt xylan source to attain the optimal culture conditions and to produce a higher yield of xylanase. The optimal culture condition was 45oC and pH 7. SM03 was identified as Bacillus(spp). SM03 was a gram positive-rod shape bacteria with spore formation and motility. The optimal xylanase activity was 0.401 IU/ml/min at 45oC and pH 7. The total protein concentration of precipitated xylanase and dialyzed xylan were 5.0mg/ml and 3.15mg/ml respectively. Precipitated xylanase had purified 2.3 fold with a yield of 90.7% whereas dialyzed xylanase gave 3.4 fold purification with a recovery of 57.2%. The xylanase gave a 0.0091 IU/s Vmax and 6.67mg/ml Km. Key words: Xylan, Xylanase, Isolation, Bacillus.