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

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. ABSTRAKMatlamat akhir kajian ini adalah untuk membuat inventori berasaskan sistematik moden daripada pencilan bakteria amilolisis yang menghasilkan bioplastik Poli-β-hidroksibutirat (PHB) daripada substrat kanji sagu. Identiti tiga pencilan bakteria tempatan telah diperiksa dalam kajian ini, menggunakan pendekatan polifasa. Satu set data berdasarkan ciriciri 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.

Isolasi dan Evaluasi Aktivitas Antifungi Bakteri Simbion Teripang Pasir (Holothuria scabra) dari Taman Nasional Wakatobi terhadap Malassezia furfur

“…Based on the results of profile matching of phenotypic characters between the selected symbiont bacterial isolates and the reference bacterial genus, it is known that these bacteria are similar to the two genera. Table 4 shows that the bacterial isolates DT1 and DT5 are identical to the key characters of the genus Pseudomonas, namely bacil/rod cell shape, Gram negative, motile, catalase positive and aerobic (Holt et al, 1994) while the DT4 isolate is identical to the key characters of the genus Bacillus, namely the cells form of bacil/rod, Gram positive, forms endospores, motile and catalase positive (Holt et al, 1994;Yanti et al, 2019). This indicates that DT1 and DT5 belong to the genus Pseudomonas while DT4 belongs to the genus Bacillus.…”

“…NaCl tolerance testing was carried out by growing cultures on Nutrient Agar (NA) media to which NaCl had been added at concentrations of 0%, 5% and 7%. Identification of symbiont bacterial isolates was carried out using the profile matching method (Yanti et al, 2019). The profile matching method is carried out by matching the characters of symbiont bacterial isolates compared to the characters of the reference genus referring to Bergey's Determinative Bacteriology (Holt et al, 1994).…”

Isolation and identification of Polyhydroxybutyrate (PHB)-producing microorganisms from various sources in North Sinai

“…It is more environmentally friendly than petroleum (Luengo et al, 2013;Thapa et al, 2018). PHB generated by microbes could be used as a plastic raw material (Yanti et al, 2019). PHB is a thermoplastic polyester with biodegradable and biocompatible features as well as physical attributes similar to polypropylene (Yanti et al, 2021).…”