Sains Malaysiana 47(3)(2018): 481–488

http://dx.doi.org/10.17576/jsm-2018-4703-07

Bioconversion of Biodiesel-derived Crude Glycerol to 1,3-Dihydroxyacetone by a Potential Acetic Acid Bacteria

(Biopenukaran Gliserol Mentah Janaan Biodiesel kepada 1,3-Dihidroksiaseton oleh Potensi Bakteria Asid Asetik)

VARAVUT TANAMOOL¹, PIYOROT HONGSACHART² & WICHAI SOEMPHOL^2*

¹Chemistry Program, Faculty of Science and Technology, Nakhon Ratchasima Rajabhat University

Nakhon Ratchasima, 30000, Thailand

²Faculty of Applied Sciences and Engineering, Khon Kaen University, Nong Khai Campus, Nong Khai, 43000, Thailand

Diserahkan: 7 Oktober 2015/Diterima: 9 Oktober 2017

ABSTRACT

Acetic acid bacteria (AAB) isolated from natural resources and fermented plant beverages were screened to produce 1,3-dihydroxyacetone (DHA) from non-detoxified crude glycerol. Among them, the isolate NKC115 was identified as Gluconobacter frateurii and produced the highest amounts of DHA. Subsequently, the effects of growth-medium conditions (initial pH, crude glycerol concentration and nitrogen sources) on growth and DHA-production capability were examined. The results showed that the crude glycerol concentration increase to above 100 g/L suppressed growth and DHA production. The highest amount of DHA obtained was 27.50 g/L, from an initial crude glycerol concentration of 100 g/L. Meanwhile, an initial pH of 5.5-7.5 in the YPGc medium did not significantly affect the bacterial growth and DHA production. The optimal nitrogen source was peptone, with DHA production at 34.70 g/L. Furthermore, overexpression of the nhaK₂ gene encoding for the Na⁺(K⁺)/H⁺ antiporter from Acetobactor tropicalis SKU1100 in G. frateurii NKC115 improved growth and increased the accumulation of DHA (37.25 g/L) from an initial crude glycerol concentration of 20%. These results indicated that the expression of this antiporter might maintain an optimal intracellular pH and concentration of Na⁺ or K⁺, leading to the cells’ ability to tolerate high concentrations of crude glycerol.

Keywords: Acetic acid bacteria; biodiesel; crude glycerol; dihydroxyacetone

ABSTRAK

Bakteria asid asetik (AAB) yang dipencilkan daripada sumber alam dan minuman fermentasi berasaskan tumbuhan telah ditapis untuk menghasilkan 1,3-dihidroksiaseton (DHA) daripada gliserol mentah yang belum disingkirkan toksiknya. Antara mereka, pencilan NKC115 telah dikenal pasti sebagai Gluconobacter frateurii dan menghasilkan jumlah DHA yang tertinggi. Seterusnya, kesan keadaan medium pertumbuhan (pH pemula, kepekatan gliserol mentah dan sumber nitrogen) terhadap pertumbuhan dan kemampuan penghasilan DHA telah dikaji. Keputusan menunjukkan kepekatan gliserol mentah bertambah melebihi 100 g/L pertumbuhan kawalan dan penghasilan DHA. Jumlah DHA tertinggi diperoleh adalah 27.50 g/L, daripada kepekatan gliserol mentah 100 g/L. Sementara itu, pH pemula 5.5-7.5 di dalam medium YPGc tidak mempengaruhi pertumbuhan bakteria dan penghasilan DHA dengan nyata. Sumber optimum nitrogen ialah pepton dengan penghasilan DHA pada 34.70 g/L. Tambahan pula, expresi melampau daripada gen pengekodan nhaK² untuk antipengangkat Na⁺(K⁺)/H⁺ daripada Acetobactor tropicalis SKU1100 dalam G. frateurii NKC115 memperbaiki pertumbuhan dan meningkatkan pengumpulan DHA (37.25 g/L) daripada kepekatan gliserol mentah pemula sebanyak 20%. Keputusan ini menunjukkan ekspresi antipengangkat berkemungkinan mengekalkan pH intrasel yang optimum dan kepekatan Na⁺ atau K⁺, menyebabkan kemampuan sel untuk menerima kepekatan gliserol mentah yang tinggi.

Kata kunci: Bakteria asid asetik; biodiesel; dihidroksiaseton; gliserol kasar

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*Pengarang untuk surat-menyurat; email: wichso@kku.ac.th