WJET  Vol.3 No.3 B , August 2015
Application of GFPuv Labeled Cronobacter sakazakii for Evaluation of Its Survival during Cornstarch Processing
Abstract: Cronobacter sakazakii is an emerging pathogen that can cause diseases for several infant groups. These bacteria were contaminated in foods, clinical utensils, and environments. In Indonesia, C. sakazakii has been isolated from powdered infant formulas, weaning foods, and other dried foods such as cornstarch. The objective of this research is to trace survival of C. sakazakii during cornstarch production step using its mutant. Mutant was constructed by inserting Green Fluorescent Protein plasmid inside to the bacterial cell that appeared green fluorescent colonies under UV observation. The presence of C. sakazakii during processing was conducted by artificial contamination. This research consists of three steps, i.e. determination of the suitable enumeration method of C. sakazakii’s mutant, cornstarch production from yellow corn, and survival analysis of C. sakazakii during endosperm soaking and cornstarch drying. The suitable enumeration method was surface plating method on TSA-ampicillin medium combining with UV light application because of ineffectiveness of ampicillin inhibition for growth of yeasts and molds. The cornstarch produced in laboratory has the same properties with commercial cornstarch in parameters of moisture content, density, and starch granule structure. The yield of cornstarch final product was 48.90% (dry whole kernel-based). C. sakazakii cannot survive in 48 hours soaking process at 52?C and 24 hours drying process at 50?C that is applied during cornstarch production.
Cite this paper: Nurjanah, S. , Sulistyanti, S. , Dewanti-Hariyadi, R. (2015) Application of GFPuv Labeled Cronobacter sakazakii for Evaluation of Its Survival during Cornstarch Processing. World Journal of Engineering and Technology, 3, 1-6. doi: 10.4236/wjet.2015.33B001.

[1]   van Acker, J., de Smet, F., Muyldermans, G., Bouhgatef, A., Naessens, A. and Lauwers, S. (2001) Outbreaks of Necrotizing Enterocolitis Associated with Enterobacter sakazakii in Powdered Milk Formula. Journal of Clinical Microbiology, 39, 293-297.

[2]   Estuningsih, S., Kress, C., Hassan, A.A., Akineden, ?., Schneider, E. and Usleber, E. (2006) Enterobacteriaceae in Dehydrated Powdered Infant Formula Manufactured in Indonesia and Malaysia. Journal of Food Protection, 69, 3013-3017.

[3]   Meutia, Y.R., Dewanti-Hariyadi, R. and Estuningsih, S. (2008) Characterization of 16S rRNA Gene of Enterobacter sakazakii Isolated from Powdered Infant Formula. In: Indonesian Association of Food Technologists, Eds., Abstracts: Annual Seminar of Indonesian Association of Food Technologists, 14-16 October 2008, Palembang.

[4]   Gitapratiwi, D., Dewanti-Hariyadi, R. and Hidayat, S.H. (2012) Genetic Relatedness of Cronobacter spp. (Enterobacter sakazakii) Isolated from Dried Food Products in Indonesia. International Food Research Journal, 19, 1745-1749.

[5]   Hamdani, F.W. (2012) Evaluation of Genetic Diversity of Local Isolates of Cronobacter spp. (Enterobacter sakazakii) Isolated from Dried Food. Thesis, Bogor Agricultural University, Bogor.

[6]   Food and Agriculture Organization-World Health Organization (FAO-WHO) (2004) Enterobacter sakazakii and Other Microorganisms in Powdered Infant FORMULA. Meeting Report, MRA Series 6, WHO, Geneva.

[7]   Dewanti-Hariyadi, R., Gitapratiwi, D., Meutia, Y.R., Hidayat, S.H. and Nurjanah, S. (2010) Isolation of Enterobacter sakazakii (Cronobacter spp.) from Powdered Infant Formula and Other Dried Foods Obtained from Bogor Area. International Seminar on Current Issues and Challenges in Food Safety: Science-Based Approach for Food Safety Management, SEAFAST Center, Bogor.

[8]   Fiegen, M. (2010) Untersuchungen zum Vorkommen und zur Tenazit?t von Cronobacter spp. Dissertation, Universit?t Hamburg Fachbereich Chemie, Hamburg.

[9]   Nurjanah, S., Dewanti-Hariyadi, R., Estuningsih, E. and Suhartono, M.T. (2014) Stability and Growth Characteristics of GFPuv-Labeled Cronobacter sakazakii Isolated from Foods. Food Science and Biotechnology, 23, 1491-1496.

[10]   Rahmawati, Dewanti-Hariyadi, R., Hariyadi, P., Fardiaz, D. and Richana, N. (2013) Isolation and Identification of Microorganisms during Spontaneous Fermentation of Maize. Jurnal Teknologi dan Industri Pangan, 24, 33-39.

[11]   Johnson, L.A. and May, J.B. (2013) Wet Milling: The Basis for Corn Bio Refineries. Chapter 12. In: White, P.J. and Johnson, L.A., Eds., Corn: Chemistry and Technology, American Asso-ciation of Cereal Chemical, Inc., St. Paul, 449- 494.

[12]   Bacteriological Analytical Manual (BAM) (2001) Chapter 3: Aerobic Plate Count.

[13]   Sharma, S.K., Singh, L. and Singh, S. (2013) Comparative Study between Penicillin and Ampicillin. Scholars Journal of Applied Medical Sciences, 1, 291-294.

[14]   Hartanti, A.W. (2007) Screening of Lactic Bacteria that Potential as Probiotic Isolated from Breast Milk. Undergraduate thesis, Bogor Agricultural University, Bogor.

[15]   Hummel, A.S., Hertel, C., Holzapfel, W.H. and Franz, C.M.A.P. (2007) Antibiotic Resistances of Starter and Probiotic Strains of Lactic Acid Bacteria. Applied and Environmental Microbiology, 73, 730-739.

[16]   Haddix, P.L., Paulsen, E.T. and Werner, T.F. (2000) Measurement of Mutation to Antibiotic Resistance: Ampicillin Resistance in Serratia marcescens. Bioscience, 26, 17-21.

[17]   Dever, L.A. and Dermody, T.S. (1991) Mechanisms of Bacterial Resistance to Antibiotics. Archives of Internal Medicine, 151, 886-895.

[18]   Iversen, C. and Forsythe, S. (2003) Risk Pro-file of Enterobacter sakazakii, an Emergent Pathogen Associated with Infant Milk Formula. Trends in Food Science & Technology, 14, 443-454.

[19]   Nazarowec-White, M. and Farber, J.M. (1997) Thermal Resistance of Enterobacter sakazakii in Reconstituted Dried-Infant Formula. Letters in Applied Microbiology, 24, 9-13.

[20]   Penna, T.C.V., Ishii, M., Junior, A.P. and Cholewa, O. (2004) Thermal Stability of Recombinant Green Fluorescent Protein (GFPuv) at Various pH Values. Applied Biochemistry and Biotechnology, 114, 469-483.