Karakterisasi Fungsi dan Peran Domain Enzim  Sucrose Phosphate Synthase  Tebu Melalui Studi Delesi Mutagenesis

Widhi Dyah Sawitri; Bambang Sugiharto

doi:10.22146/veg.81130

Karakterisasi Fungsi dan Peran Domain Enzim Sucrose Phosphate Synthase Tebu Melalui Studi Delesi Mutagenesis

https://doi.org/10.22146/veg.81130

Widhi Dyah Sawitri^(1*), Bambang Sugiharto⁽²⁾

(1) Fakultas Pertanian, Universitas Gadjah Mada
(2) Jurusan Biologi, FMIPA, Universitas Jember
(*) Corresponding Author

Abstract

Sucrose phosphate synthase (SPS) berperan penting untuk biosintesis sukrosa pada tanaman. SPS tanaman dibagi menjadi beberapa domain diantaranya bagian domian N-terminal, domain C-terminal, dan domain glycosyltransferase (GT). Namun, fungsi domain N-terminal dan C-terminal masih belum jelas. Oleh karenanya, dilakukan studi karakterisasi fungsi masing-masing domain SPS melalui penghilangan domain N-terminal (∆N), C-terminal (∆C), dan kedua domain tersebut (∆NC). Mutan ∆N, ∆C, dan ∆NC diekspresikan melalui sistem rekombinan E. coli dan dilakukan purifikasi protein secara parsial untuk menghilangkan sebagian protein kontaminan. Protein rekombinan mutan dikonfirmasi ekspresinya melalui teknik western blot dan dilakukan uji aktivitas enzim. Hasil studi menunjukkan bahwa ekspresi dan aktivitas enzim mutan ∆N meningkat. Sebaliknya, pada mutan ∆C dan ∆NC terjadi penurunan aktivitas enzim, bahkan mendekati kondisi tidak aktif. Hal ini mengindikasikan bahwa bagian domain C-terminal SPS tanaman memiliki peran yang sangat krusial untuk sintesis sukrosa. Melalui studi ini dapat diketahui bahwa domain N-terminal diindikasikan berperan dalam regulasi alosterik enzim, sedangkan domain C-terminal diasumsikan berkaitan dengan reaksi katalitik enzim.

Keywords

sucrose phosphate synthase (SPS); mutasi delesi; metabolisme sukrosa; tebu

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DOI: https://doi.org/10.22146/veg.81130