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Heterologous expression and characterization of thermostable lipase (Lk1) in Pichia pastoris GS115
Nurul Furqan B.R.a, Akhmalokaa,b
a Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Indonesia
b Department of Chemistry, Faculty of Science and Computer, Universitas Pertamina, Indonesia
[vc_row][vc_column][vc_row_inner][vc_column_inner][vc_separator css=”.vc_custom_1624529070653{padding-top: 30px !important;padding-bottom: 30px !important;}”][/vc_column_inner][/vc_row_inner][vc_row_inner layout=”boxed”][vc_column_inner width=”3/4″ css=”.vc_custom_1624695412187{border-right-width: 1px !important;border-right-color: #dddddd !important;border-right-style: solid !important;border-radius: 1px !important;}”][vc_empty_space][megatron_heading title=”Abstract” size=”size-sm” text_align=”text-left”][vc_column_text]© 2019A gene encoding thermostable lipase namely LK1 was successfully sub-cloned into expression vector pPICZαA and integrated into a chromosomal fungal host, Pichia pastoris GS115. The recombinant mut+ clones selected on zeocine-YPD plates were heterologically expressed into Pichia pastoris GS115 using 1% methanol as inducer at 30°C during 120 h in BMMY medium. The protein was expressed at molecular mass 35.5 kDa following SDS-PAGE analysis. The enzyme still showed lipase activity on pNP as substrate. Thermostable lipase was purified by affinity chromatography using Ni-NTA column with a purification fold of 11.6 and yield of 31.75%. Further characterization of the enzyme showed that the enzyme has highest spesicificity on pNP-laurate as substrate with optimum temperature at 60 °C, pH 8. In addition, the enzyme still maintained the activity up to 4 h in incubation at 60 °C and maintained 50% activity at 3 h incubation. All of the above data suggested that the enzyme is an alkaline tolerance and thermostable lipase.[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Author keywords” size=”size-sm” text_align=”text-left”][vc_column_text][/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Indexed keywords” size=”size-sm” text_align=”text-left”][vc_column_text]Heterologous expression,Ni-NTA column chromatography,Pichia pastoris GS115,Thermostable lipase[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Funding details” size=”size-sm” text_align=”text-left”][vc_column_text]We would like to thank to P3MI research program of ITB and LPDP (scholarship to BRNF) to make this research possible to be carried out.[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”DOI” size=”size-sm” text_align=”text-left”][vc_column_text]https://doi.org/10.1016/j.bcab.2019.101448[/vc_column_text][/vc_column_inner][vc_column_inner width=”1/4″][vc_column_text]Widget Plumx[/vc_column_text][/vc_column_inner][/vc_row_inner][/vc_column][/vc_row][vc_row][vc_column][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][/vc_column][/vc_row]