Functional characterization of the sucrose isomerase responsible for trehalulose production in plant-associated Pectobacterium species |
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| Institution: | 1. Graduate School of Biotechnology, and Institute of Life Science and Resources, Kyung Hee University, Yongin 446-701, Republic of Korea;2. Department of Plant Medicine, Suncheon National University, Suncheon 540-950, Republic of Korea;3. Department of Biology, Suncheon National University, Suncheon 540-950, Republic of Korea;4. Microbial Safety Division, National Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Republic of Korea;5. Department of Horticultural Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea;1. Department of Medicinal Chemistry, Institute of Pharmacology, Polish Academy of Sciences, Smetna 12, Kraków 31-343, Poland;2. Faculty of Chemistry, Jagiellonian University, R. Ingardena 3, Kraków 30-060, Poland;3. Faculty of Mathematics and Computer Science, Jagiellonian University, S. Lojasiewicza 6, Kraków 30-348, Poland;1. Department of Pre-Clinical Science (Biochemistry), Faculty of Medicine, Thammasat University, Pathumthani 12120, Thailand;2. Department of Biochemistry, Phramongkutklao College of Medicine, Phramongkutklao Hospital, Bangkok 10400, Thailand;3. Protein Research Laboratory, Faculty of Medicine, Thammasat University, Pathumthani 12120, Thailand;4. Department of Biology, Graduate School of Science, Osaka City University, Osaka 558-8585, Japan;5. Starch and Cyclodextrin Research Unit, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand;1. Department of Nutrition and Dietetics, School of Health Sciences, International Medical University, Bukit Jalil, 57000 Kuala Lumpur, Malaysia;2. Jeffrey Cheah School of Medicine and Health Sciences, Monash University Malaysia, Jalan Lagoon Selatan, 47500 Bandar Sunway, Selangor Darul Ehsan, Malaysia |
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| Abstract: | Fifty-three plant-associated microorganisms were investigated for their ability to convert sucrose to its isomers. These microorganisms included one Dickeya zeae isolate and 7 Enterobacter, 3 Pantoea, and 43 Pectobacterium species. Eleven out of the 53 strains (21%) showed the ability to transform sucrose to isomaltulose and trehalulose. Among those, Pectobacterium carotovorum KKH 3-1 showed the highest bioconversion yield (97.4%) from sucrose to its isomers. In this strain, the addition of up to 14% sucrose in the medium enhanced sucrose isomerase (SIase) production. The SIase activity at 14% sucrose (47.6 U/mg dcw) was about 3.6-fold higher than that of the negative control (13.3 U/mg dcw at 0% sucrose). The gene encoding SIase, which is comprised a 1776 bp open reading frame (ORF) encoding 591 amino acids, was cloned from P. carotovorum KKH 3-1 and expressed in Escherichia coli. The recombinant SIase (PCSI) was shown to have optimum activity at pH 6.0 and 40 °C. The reaction temperature significantly affected the ratio of sucrose isomers produced by PCSI. The amount of trehalulose increased from 47.5% to 79.1% as temperature was lowered from 50 °C to 30 °C, implying that SIase activity can be controlled by reaction temperature. |
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| Keywords: | Isomaltulose Sucrose Sucrose isomerase Trehalulose |
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