Sains Malaysiana 48(12)(2019): 2663–2673

http://dx.doi.org/10.17576/jsm-2019-4812-07

 

Polyphasic Identification of Amylolytic Bacteria Producing Bioplastic Poly-β-hydroxybutyrate (PHB)

(Pengenalpastian Polifasa Bakteria Amilolisis Menghasilkan Bioplastik Poli-ß-hidroksibutirat (PHB))

 

NUR ARFA YANTI1*, L. SEMBIRING2, S. MARGINO3, NURHAYANI H. MUHIDDIN4, SITTI WIRDHANA AHMAD1 & ARDIANSYAH1

 

1Department of Biology, Faculty of Mathematics and Natural Science, Halu Oleo University, Kendari Southeast Sulawesi, Indonesia

 

2Faculty of Biology, Gadjah Mada University, Yogyakarta, Indonesia

 

3Faculty of Agricultural, Gadjah Mada University, Yogyakarta, Indonesia

 

4Faculty of Mathematics and Natural Science, Makassar State University, Makassar, South Sulawesi, Indonesia

 

Received: 27 April 2019/Accepted: 25 September 2019

 

ABSTRACT

The final goal of this study is to make a modern systematic-based inventory of amylolytic bacterial isolates producing of bioplastic Poly-β-hydroxybutyrate (PHB) from sago starch substrate. The identity of three local bacterial isolates was examined in this study, using a polyphasic approach. A data set based on phenotypic characteristics, namely morphological, physiological, biochemical and chemical character, namely whole cells protein profiles using SDS-PAGE method, together with phylogenetic studies based on 16S rRNA sequences was used to identified by polyphasic approach. Phenotypic characteristics of 3 local bacterial isolates and 4 reference strains to members of genus Bacillus was analyzed by numerical analysis using MVSP 3,1 program to determine the value of similarity. Based on the preliminary characterization of the profile matching method showed that the three isolates of bacteria producing PHB namely PSA10, PPK5 and PPK6 are members of the genus Bacillus. The results of numerical analysis based on phenotypic characteristic and chemical character of the three bacterial isolates producing PHB with reference strains showed that the PSA10 isolate bacterial identical with Bacillus megaterium, PPK5 isolate identical with Bacillus subtilis and PPK6 isolate identical with Bacillus cereus, and these results also support by the molecular phylogenetic analysis. Therefore, the polyphasic taxonomy is an effective approach to uncover the identity of the novel bacterial isolates.

Keywords: Amylolytic bacteria; bioplastic; numeric-phenetic; PHB; polyphasic

 

ABSTRAK

Matlamat akhir kajian ini adalah untuk membuat inventori berasaskan sistematik moden daripada pencilan bakteria amilolisis yang menghasilkan bioplastik Poli-ß-hidroksibutirat (PHB) daripada substrat kanji sagu. Identiti tiga pencilan bakteria tempatan telah diperiksa dalam kajian ini, menggunakan pendekatan polifasa. Satu set data berdasarkan ciri-ciri fenotip, iaitu sifat morfologi, fisiologi, biokimia dan kimia, iaitu profil protein sel keseluruhan menggunakan kaedah SDS-PAGE, bersama-sama dengan kajian filogenetik berdasarkan urutan 16S rRNA digunakan untuk mengenal pasti dengan pendekatan polifasa. Ciri-ciri fenotip 3 pencilan bakteria tempatan dan 4 rujukan kepada ahli genus Bacillus dianalisis dengan analisis berangka menggunakan MVSP 3,1 program untuk menentukan nilai keserupaan. Berdasarkan pencirian permulaan kaedah pencocokan profil menunjukkan bahawa tiga pencilan bakteria yang menghasilkan PHB iaitu PSA10, PPK5 dan PPK6 adalah ahli genus Bacillus. Hasil analisis berangka berdasarkan pencirian fenotip dan sifat kimia ketiga pencilan bakteria yang menghasilkan PHB dengan strain rujukan menunjukkan bahawa PSA10 seiras dengan Bacillus megaterium, PPK5 seiras dengan Bacillus subtilis dan PPK6 seiras dengan Bacillus cereus, dan keputusan ini juga disokong oleh analisis filogenetik molekul. Oleh itu, taksonomi polifasa adalah pendekatan yang berkesan untuk mendedahkan identiti novel pencilan bakteria.

Kata kunci: Bakteria amilolisis; bioplastik; numerik-fenetik; PHB; polifasa

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*Corresponding author; email: nur.yanti@uho.ac.id

 

 

 

 

 

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