ANALISIS SEKUEN POLIKETIDA SINTASE DOMAIN KETOSINTASE PADA BAKTERI ENDOFIT AKAR AGERATUM CONYZOIDES L

Saputra, Susadi Nario (2013) ANALISIS SEKUEN POLIKETIDA SINTASE DOMAIN KETOSINTASE PADA BAKTERI ENDOFIT AKAR AGERATUM CONYZOIDES L. S1 thesis, Universitas Pendidikan Indonesia.

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Abstract

Polyketida synthase (PKS) merupakan kelompok enzim multi kompleks yang bertanggungjawab dalam proses biosintesis berbagai macam senyawa poliketida. Poliketida sintase mempunyai beberapa modul yang terdiri atas beberapa domain dengan fungsi yang berbeda. Ketosintase (KS) merupakan salah satu domain yang diperlukan untuk proses elongasi atau pemanjangan rantai poliketida. Ketersediaan dan diversitas domain ketosintase di alam penting dalam usaha penemuan obatan – obatan baru. Telah dilakukan kloning gen ketosintase bakteri endofit akar Ageratum conyzoides L ke dalam vektor pGEMT – Easy. Gen ketosintase diperoleh dari bakteri endofit akar Ageratum conyzoides dengan metode isolasi DNA secara lansung melaui proses enrichment. Primer DKF/DKR dan HGLF/HGLR digunakan untuk mengamplifikasi domain ketosintase secara lansung dari berbagai sampel lingkungan. Amplikon diklon kedalam plasmid pGEMT – Easy dan ditransformasikan kedalam bakteri Escherichia coli DH5α. Seleksi transforman dilakukan dengan metode seleksi biru putih. Efesiensi transformasi diperoleh sebesar 6.68 X 103 CFU/µg untuk gen ketosintase heterokyst glicolipid dan 3.12 X 103 CFU/µg untuk gen ketosintase non heterokyst glicolipid. Hasil analisis filogenetik menunjukkan bahwa telah berhasil teridentifikasi domain ketosintase dari berbagai kelompok bakteri meliputi, Actinobacteria, Cyanobacteria, Proteobacteria, dan beberapa mikroorganisme yang belum teridentifikasi. Keyword : Polyketida sintase, Ketosntase, Ageratum conyzoides, Kloning Polyketide synthase (PKS) are known to produce a class of medically and industrially include a broad range of bioactive compound such antibiotics. PKSs are multifungsional enzyme that organized into module and consist of several domain. KS domain is responsible for the condensation of an extender unit onto the growing polyketide chain during polyketide biosynthesis. The diversity of modular polyketide synthase (PKS) gene in root of Ageratum conyzoides were studied by sequencing analysis than before clone in DH5α bacteria. Primer for the amplification of PKS domain were DKF and DKR for non heterocyst glicolipid and HGLF and HGLR for heterocyst glikolipid. The eficience of transformation 6.68 X 103 CFU/µg for heterokyst glicolipid gene and 3.12 X 103 CFU/µg for heterokyst glicolipid gene. Phylogenetic analysis of 20 amino acid (AA) sequences indicates that the identified ketosynthase (KS) domains were clustered with those from diverse bacterial groups, including Cyanobacteria, Proteobacteria, Actinobacteria and some unidentified microorganisms. Phylogenetic result show DK10 DK8 and DK3 include protobacteria group, DK6 DK7 and DK9 include Cyanobacteria, DK4 DK7 include Actinobacteria group. Phylogentic analysis of heterocyst glycolipid biosynthetic ketoshynthase region with respect to diverse range of ketosynthase domain, including type I and II Keyword : Polyketide sinthase, Ketosynthase, Ageratum conyzoides, Cloning

Item Type: Skripsi,Tesis,Disertasi (S1)
Subjects: Universitas Pendidikan Indonesia > Fakultas Pendidikan Matematika dan Ilmu Pengetahuan Alam > Jurusan Pendidikan Biologi > Program Studi Pendidikan Biologi
Divisions: Fakultas Pendidikan Matematika dan Ilmu Pengetahuan Alam > Jurusan Pendidikan Biologi > Program Studi Pendidikan Biologi
Depositing User: DAM STAF Editor
Date Deposited: 11 Nov 2013 04:32
Last Modified: 11 Nov 2013 04:32
URI: http://repository.upi.edu/id/eprint/3004

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