Browse > Article
http://dx.doi.org/10.22805/JIT.2019.39.1.015

Characterization of the Starch Degradation Activity of recombinant glucoamylase from Extremophile Deinococcus geothermalis  

Jang, Seung-Won (ForBioKorea Co., Ltd.)
Kwon, Deok-Ho (Department of Bioengineering and Technology, Kangwon National University)
Park, Jae-Bum (Department of Bioengineering and Technology, Kangwon National University)
Jung, Jong-Hyun (Research Division for Biotechnology, Korea Atomic Energy Research Institute)
Ha, Suk-Jin (Department of Bioengineering and Technology, Kangwon National University)
Publication Information
Journal of Industrial Technology / v.39, no.1, 2019 , pp. 15-19 More about this Journal
Abstract
This work focused on characterization of the starch degradation activity from extremophile strain Deinococcus geothermalis. Glucoamylase gene from D. geothermalis was cloned and overexpressed by pET-21a vector using E. coli BL21 (DE3). In order to characterize starch degrading activity of recombinant glucoamylase, enzyme was purified using HisPur Ni-NTA column. The recombinant glucoamylase from D. geothermalis exhibited the optimum temperature as $45^{\circ}C$ for starch degradation activity. And highly acido-stable starch degrading activity was shown at pH 2. For further optimization of starch degrading activity with metal ion, various metal ions ($AgCl_2$, $HgCl_2$, $MnSO_4{\cdot}4H_2O$, $CoCl_2{\cdot}6H_2O$, $MgSO_4$, $ZnSO_4{\cdot}7H_2O$, $K_2SO_4$, $FeCl_2{\cdot}4H_2O$, NaCl, or $CuSO_4$) were added for enzyme reaction. As results, it was found that $FeCl_2{\cdot}4H_2O$ or $MnSO_4{\cdot}4H_2O$ addition resulted in 17% and 9% improved starch degrading activity, respectively. The recombinant glucoamylase from D. geothermalis might be used for simultaneous saccharification and fermentation (SSF) process at high acidic conditions.
Keywords
Deinococcus geothermalis; glucoamylase; pET-21a vector; simultaneoussaccharification and fermentation;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Bialas W., Szymanowska D., Grajek W., 2010, Fuel ethanol production from granular corn starch using Saccharomyces cerevisiae in a long term repeated SSF process with full stillage recycling, Bioresource technology, 101 3126-3131.   DOI
2 Keith C., 2006, Economic Issues related to bio fuels; a written testimony for field hearing US senate committee on agriculture, rular development, and related agencies.
3 Ueda S., 1981, Fungal glucoamylases and raw starch digestion, Trends in Biochemical Sciences, 6 89-90.   DOI
4 Shigechi H., Koh J., Fujita Y., Matsumoto T., Bito Y., Ueda M., Satoh E., Fukuda H., Kondo A., 2004, Direct production of ethanol from raw corn starch via fermentation by use of a novel surface-engineered yeast strain codisplaying glucoamylase and ${\alpha}$-amylase. Appl. Environ. Microbiol., 70 5037-5040.   DOI
5 Aggarwal N., Niga P., Singh D., Yadav B., 2001, Process optimization for the production of sugar for the bioethanol industry from Tapioca, a non-conventional source of starch, World Journal of Microbiology and Biotechnology, 17 783-787.   DOI
6 Kim S., Dale B.E., 2004, Global potential bioethanol production from wasted crops and crop residues, Biomass and bioenergy, 26 361-375.   DOI
7 Teixeira L.C., Linden J.C,. Schroeder H.A., 1999, Optimizing peracetic acid pretreatment conditions for improved simultaneous saccharification and co-fermentation (SSCF) of sugar cane bagasse to ethanol fuel, Renewable Energy, 16 1070-1073.   DOI
8 Morales-Rodriguez R., Gernaey K.V., Meyer A.S., Sin G., 2011, A mathematical model for simultaneous saccharification and co-fermentation (SSCF) of C6 and C5 sugars, inese Journal of Chemical Engineering, 19 185-191.   DOI
9 Mukerjea R., Slocum G., Mukerjea R., Robyt J.F., 2006, Significant differences in the activities of ${\alpha}$-amylases in the absence and presence of polyethylene glycol assayed on eight starches solubilized by two methods, Carbohydrate research, 341 2049-2054.   DOI
10 Huang L.P., Jin B., Lant P., Zhou J., 2005, Simultaneous saccharification and fermentation of potato starch wastewater to lactic acid by Rhizopus oryzae and Rhizopus arrhizus, Biochemical Engineering Journal, 23 265-276.   DOI
11 Kim N., Kim J.S., 2004, Acetic Acid Production from Cellulosic Biomass by Simultaneous Saccharification and Fermentation, Theories Applicat., Chem., Eng10, 1546-1549.
12 Oh K.W., Kim M.J., Kim H.Y., Kim B.Y., Baik M.Y., Auh J.H., Park C.S., 2005, Enzymatic characterization of a maltogenic amylase from Lactobacillus gasseri ATCC 33323 expressed in Escherichia coli, FEMS microbiology letters, 252 175-181.   DOI
13 Jang S.W., Kim J.S., Park J.B., Jung J.H., Park C.S., Shin W.C., Ha S.J., 2015, Characterization of the starch degradation activity from newly isolated Rhizopus oryzae WCS-1 and mixed cultures with Saccharomyces cerevisiae for efficient ethanol production from starch, Food science and biotechnology, 24 1805-1810.   DOI
14 Brim H., McFarlan S.C., Fredrickson J.K., Minton K.W., Zhai M., Wackett L.P., Daly M.J., 2000, Engineering Deinococcus radiodurans for metal remediation in radioactive mixed waste environments, Nature biotechnology, 18 85.   DOI
15 Panek A., Pietrow O., Synowiecki J., 2012, Characterization of glucoamylase immobilized on magnetic nanoparticles, Starch-Starke, 64 1003-1008.   DOI
16 Ferreira A.C., Nobre M.F., Rainey F.A., Silva M.T., Wait R., Burghardt J., Chung A.P., Da Costa M.S., 1997, Deinococcus geothermalis sp. nov. and Deinococcus murrayi sp. nov., two extremely radiation-resistant and slightly thermophilic species from hot springs, International Journal of Systematic and Evolutionary Microbiology, 47 939-947.
17 Makarova K.S., Aravind L., Wolf Y.I., Tatusov R.L., Minton K.W., Koonin E.V., Daly M.J., 2001, Genome of the extremely radiation-resistant bacterium Deinococcus radiodurans viewed from the perspective of comparative genomics, Microbiol. Mol. Biol. Rev.65 44-79.   DOI