• Journal of Microbiology and Biotechnology
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• v.14 no.4
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• pp.686-692
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• 2004

To develop the enzyme-linked immunosorbent assay (ELISA) for the analysis of N-acetylehitooligosaccharides (NACOS) and chitooligosaccharides (COS), specific antibodies (Abs) were produced, and their properties were compared. N-acetylehitohexaose (NACOS6), chitohexaose (COS6), and COS mixture (COSM) conjugated to bovine serum albumin (BSA) were used to immunize rabbits. By the use of specific Abs and NACOS6-horseradish peroxidase (HRP), COS6-HRP, and COSM-HRP conjugates, competitive direct ELISA (cdELISA) was developed. The detection limits of NACOS6 by the anti-NACOS6 Ab and COS6 by the anti-COS6 and the anti-COSM Abs in the cdELISAs were about 0.2, 2, ana 2 ng/ml (ppb), respectively. In the cdELISA, the anti-NACOS6 Ab was found to recognize NACOS3-NACOS6, but not N-acetyl-D-glucosamine (GlcNAc), NACOS2, and COSs; the anti-COS6 Ab recognized COS2-COS6 and COSM, but not glucosamine (GlcN) and NACOSs. The recognition pattern of the anti-COSM Ab was almost the same as that of the anti- COS6 Ab, except that the former recognized COS2 and COS3 slightly better than the latter.

• Journal of Microbiology and Biotechnology
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• v.14 no.4
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• pp.877-880
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• 2004

In this study, we determined the content of chitooligosaccharides (COS) in Korean commercial salt-fermented shrimps by competitive direct enzyme-linked immunosorbent assays (cdELISAs), using anti-COS mixture (COSM) antibody and COSM horseradish peroxidase (HRP) conjugate. When COS6 was spiked into salt-fermented shrimps at the level of $10-300\mu{g/g,}$ the average recovery was $120\pm19%$ ($mean\pmS.D.$). The COS contents of the 92 samples of Korean commercial salt-fermented shrimps collected during February 2000 and August 2002 were $36.3\pm20.7\mug$ COS6 equivalent/g (expressed as "$\mug/g$" hereafter). Among the samples, the COS contents of yuk-jeot ( $40.3 \pm 22.5 \mug/g, n=27$) and buksaewoojeot ($40.2 \pm 21.6 \mug/g, n=5$) were higher than the others. The COS contents of salt-fermented shrimps produced at Gwangcheon ($47.1 \pm 20.7 \mug/g, n=18$) and Gomso ($44.1 \pm 21.8 \mug/g, n=6$) areas were higher than those produced at the other areas. This is the first report to determine COS of salt-fermented shrimps by cdELISA.

• Journal of Microbiology and Biotechnology
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• v.14 no.2
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• pp.256-261
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• 2004

In order to detect chitooligosaccharides (COS) in soybean paste, tandem immunoaffinity chromatography and enzyme-linked immunosorbent assay (ELISA) were developed. Polyclonal anti-chitooligosaccharides mixture (CaSM) antibody specific to COSM was attached to Sepharose gel for initial sample cleanup and concentration of COS in soybean paste. COS was eluted and quantified by competitive direct ELISA (cdELISA). Average ELISA recoveries from the column using binding buffer spiked with COSM at levels of 0.5, 2.0, 5.0, and $10.0\mu$g/ml were 79.8, 72.0, 77.7, and 60.6%, respectively, with a mean recovery of 72.5%. Mean inter-well and inter-assay coefficients of variation (CV) were 7.7% and 10.3%, respectively. Average recoveries from soybean paste spiked with COSM at levels of 2, 6, 20, and $60\mu$g/g were 115, 91.7, 91, and 73.3%, respectively, with a mean recovery of 92.8%. Mean inter-well and inter-assay CV were 12.9% and 16%, respectively. The COS was detected from 24 out of 25 homemade Korean soybean paste samples at an average of $14.0\mu$g/g (n, 25; range, $0-51.2 \mu$g/g) and from 13 out of 14 commercially made soybean paste samples at an average of $4.1\mu$g/g(n, 14; range, $0-18.4\mu$g/g). The tandem immunoaffinity chromatography-cdELISA that was developed in this study showed that the level of COS eluted from homemade soybean paste was higher than that of the commercially made ones. In addition, the level of COS eluted from commercially available soybean paste in Korea was higher than that of the ones in Japan.

• Korean Journal of Food Science and Technology
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• v.35 no.2
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• pp.291-296
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• 2003

Effects of Bacillus subtilis and Rhizopus oryzae on chitooligosaccharides (COS) content of doenjang (soybean paste) and kanjang (soy sauce) were investigated using kojis made with the two strains. Competitive direct enzyme-linked immunosorbent assay (cdELISA) system using anti-COS mixture (COSM) antibody was applied for COS detection ranging from 0.001 to $1{\mu}g/mL$, and the recoveries of COSM spiked to doenjang and kanjang were 102 and 115%, respectively. Doenjang and kanjang products made with a mixture of B. subtilis and R. oryzae kojis showed COS contents of 171 and $29{\mu}g/mL$, respectively, during two-month aging period, much higher than those of Japanese and Korean commercial ones.

• Journal of Microbiology and Biotechnology
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• v.20 no.7
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• pp.1114-1120
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• 2010

A chitosan-degrading fungus, BSF114, was isolated from soil. The culture preparation showed strong chitosanolytic enzyme activity at an optimum pH of 4.0 and optimum temperature of $60^{\circ}C$ after 36-40 h fermentation. The rapid decrease in the viscosity of the chitosan solution early in the reaction suggested an endo-type cleavage of the polymeric chitosan chains. To identify the isolated fungus, molecular biological and morphological methods were used. The fungal internal transcribed spacer (ITS) region 1 was amplified, sequenced, and then compared with related sequences in the GenBank database using BLAST. The phylogenetic relationships were then analyzed, and the results showed that the fungus belongs to Aspergillus fumigatus. Morphological observations were also used to confirm the above conclusion. The chitooligosaccharides (COS) obtained through hydrolyzing the colloidal chitosan showed that A. fumigatus BSF114 is suitable for degrading chitosan and producing chitooligosaccharides on a large scale. High concentrations of the COS (1,000 and 500 ${\mu}g/ml$) significantly proliferated mice marrow cells.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.86-87
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• 2000

Chitosan (deacetylated form of chitin) possesses strong antibacterial activities such as antimicrobial effect, antifungal effect and the induction of plant defense response. Chitosan itself, however, has high molecular weight and viscosity as well as water-insolubility, These natures may restrict applications in various fields, especially in in vivo system. While the hydrolysates of chitosan, chitooligosaccharides (COS) are not only lower in the molecula. weight and viscosity, but also water-soluble. Thus, they would be expected more efficient absorption in vivo. Besides several documents have been reported antibacterial activities of COS against microorganisms (Kendra et al., 1989; Uchida et al., 1989). (omitted)

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.2
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• pp.236-243
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• 2004

Antimicrobial effect of chitosan and its oligosaccharides was examined on Vibrios sp., Edwardsiella tarda and Streptococcus sp., which are major pathogenic bacteria inducing bacterial diseases of acquacultured flounder. Chitosan oligosaccharides (COS ) were produced by enzymatic hydrolysis of chitosan in an ultrafiltration mombrane bioreactor system which was established with three membranes with different molecular weight cut-off (MWCO) 1,000, 5,000 and 10,000, and fractionated into three kinds of COS, based on their molecular weight sizes. The three kinds of COS were as follows : relatively high molecular weight COS [HMW-COS, molecular weight distribution of 7,000 to 24,000 Da〕, medium molecular weight COS 〔MMW-COS, 1,500 to 6,000 Da〕, and low molecular weight COS 〔LMW-COS, 1,000 to 1,500 Da). Chitosan and HMW-COS effectively inhibited the growths of Vibrio sp. and Streptococcus sp. and their antimicrobial activities were superior to the others with smaller molecular weights. This result suggested that antimicrobial effect of chitosan preparations extremely depend on their molecular weight sizes. Antimicrobial effect of chitosan and HMW-COS on E. tarda was improved by longer inoculation times. Scanning electron microscopy in morphological change of E. tarda treated with chitosan preparations showed that chitosan and HMW-COS bound to the cells and suppressed the growth of the cells. This observation appears to prove the fact that positive charged amines of chitosan electrostatically bind to negative charged compounds of cell walls.

• Food Science and Industry
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• v.53 no.1
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• pp.2-32
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• 2020

In recent years, significant importance has been given to chitooligosaccharides (COS) due to its potent notable biological applications. COS can be derived from chitosan which is commonly produced by partially hydrolyzed products from crustacean shells. In order to produce COS, there are several approaches including chemical and enzymatic methods which are the two most common choices. In this regard, several new methods were intended to be promoted which use the enzymatic hydrolysis with a lower cost and desired properties. Hence, the dual reactor system has gained more attention than other newly developed technologies. Enzymatic hydrolysis derived COS possesses important biological activities such as anticancer, antioxidant, anti-hypersentive, anti-dementia (Altzheimer's disease), anti-diabeties, anti-allergy, anti-inflammatory, etc. Results strongly suggest that properties of COS can be potential materials for nutraceutical, pharmaceutical, and cosmeceutical product development.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.311-318
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• 2010

Antimicrobial finishing of textiles has been found to be an economical way to prevent (or treat) skin disorders. Hence, this research effort was aimed at elucidating the relationship between the molecular weight (MW) of chitosan and its antimicrobial activity upon six dermal reference microorganisms, as well as the influence of the interactions with cotton fabrics on said activity. Using 3 chitosans with different MWs, as well as two chitooligosaccharide (COS) mixtures, a relevant antimicrobial effect was observed by 24 h for the six microorganisms tested; it was apparent that the antimicrobial effect is strongly dependent on the type of target microorganism and on the MW of chitosan - being higher for lower MW in the case of E. coli, K. pneumoniae, and P. aeruginosa, and the reverse in the case of both Gram-positive bacteria. Furthermore, a strong antifungal effect was detectable upon C. albicans, resembling the action over Gram-positive bacteria. Interactions with cotton fabric resulted in a loss of COS activity when compared with cultured media, relative to the effect over Gram-negative bacteria. However, no significant differences for the efficacy of all the 5 compounds were observed by 4 h. The three chitosans possessed a higher antimicrobial activity when impregnated onto the fabric, and presented a similar effect on both Gram-positive bacteria and yeast, in either matrix. Pseudomonas aeruginosa showed to be the most resistant microorganism to all five compounds.