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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)
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
Diserahkan: 27
April 2019/Diterima: 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;
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*Pengarang untuk surat-menyurat; email: nur.yanti@uho.ac.id
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