- Email: [email protected]
Fractional factorial design and central composite design for CO2 mitigation using Senedesmus producto-capitatus
Abstracts / Journal of Biotechnology 136S (2008) S589–S601
total carbohydrate content is, however, 10% lower than that of the control.
References Fidalgo, J.P., Cid, A., Torres, E., Sukenik, A., Herrero, C., 1998. Effects of nitrogen source and growth phase on proximate biochemical composition, lipid classes and fatty acid profile of the marine microalgae Isochrysis galbana. Aquaculture 166 (1998), 105–116. Jeffrey, S.W., Brown, M.R., Volkman, J.K., 1994. Haptophyte as feed stocks in mariculture. In: Green, J.C., Leadbeater, B.S.C. (Eds.), The Haptophyte Algae. Clarendon Press, Oxford, pp. 287–302.
doi:10.1016/j.jbiotec.2008.07.1201 VI6-P-017 A study on sponge associated actinomycetes and isolation of potent antimicrobial compounds V. Vaishnavapriya 1,∗ , R. Venkatesan 2 , N. Manoharan 3 1
Center for Ocean research, Sathyabama University, Chennai 119, India Ocean Science and Technology for Islands, National Institute of Ocean Technology, Chennai 601302, India 3 Sathyabama University, Chennai 119, India 2
E-mail address: [email protected] (V. Vaishnavapriya). Sponges are well known to harbor diverse microbes and represent a significant source of bioactive compounds. As far as the actinomycetes are concern, they are the prime producers of antibiotics. It is strongly believed that the microbes do produce the potent secondary metabolites at the extreme stress conditions which do possess potent antimicrobial activity. The increasing occurrence of infectious disease is becoming a world wide problem. Additionally, the resistance problem demands that the renewal efforts be made to seek antimicrobial agents that are effective against pathogenic microorganisms resistant to current treatment. So this work mainly aims to isolate sponge associated actinomycetes and is targeted to extract the potent antimicrobial activity against clinical strains. The sponge extract of 60 g/concentration is very effective against the pathogenic bacterium. The Streptomycetes sp. isolated from the sponge shows the inhibitory property against the clinical strains. doi:10.1016/j.jbiotec.2008.07.1202 VI6-P-018 Comparative study of N-trimethyl chitosan (TMC) and chitooligosaccharides on anti-radical scavenging activity A. Ozhan Aytekin ∗ , Shigeru Morimura Graduate School Science and Technology, Kumamoto University, 2-391 Kurokami, Kumamoto 860-8555, Japan E-mail address: [email protected] (A.O. Aytekin). Chitosan is a biopolymer and mainly found in chitin form in crustacean shells. Chitosan, which is copolymer consisting of beta-(1 → 4)-2-acetamido-d-glucose and beta-(1 → 4)-2-amino-dglucose, units, is linear and natural cationic polysaccharide. Although chitosan is biocompatible, biodegradable and non-toxic polymer, its solution has high viscosity and it is mainly dissolved in acidic solutions. Therefore, its derivatives and oligomers are considered recently (Kurita, 2006; Kim and Rajapakse, 2005). In this study, initially N-trimethyl chitosan (TMC) were produced from commercial chitosan (400 kDa) and chitosan oligomers (1 kDa). Then char-
S595
acterization of chitosan, chitosan oligomers and TMC were done by H NMR, HPLC and viscosimeter (Amidi et al., 2006). Antiradical scavenging activity on the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical of three different degree of quaternization (DQ) of TMC and two different chitosan depending on molecular weight were investigated (Lin and Chou, 2004). The results showed that, TMC has higher antiradical activity than chitosan and its oligomers. Lower DQ and molecular weight of TMC and chitosan have higher scavenging effect (SE), separately. According to positive control, which is ascorbic acid at 1 mg/ml, 95.5% SE is observed and at the same concentration TMC, chitosan (400 kDa) and chitosan oligomers (1 kDa) ranging between 75% and 85%; 10–15%; 27–35% SE, respectively. References Amidi, M., Romeijn, S.G., Borchard, G., Junginger, H.E., Hennink, W.E., Jiskoot, W., 2006. Preparation and characterization of protein-loaded N-trimethyl chitosan nanoparticles as nasal delivery system. J. Control. Release 111, 107–116. Kurita, K., 2006. Chitin and chitosan: functional biopolymers from marine crustaceans. Mar. Biotechnol. 1–24. Kim, S.K., Rajapakse, N., 2005. Enzymatic production and biological activities of chitosan oligosaccharides (COS): a review. Carbohyd. Polym. 62, 357–368. Lin, H.Y., Chou, C.C., 2004. Antioxidative activities of water-soluble disaccharide chitosan derivatives. Food Res. Int. 37 (9), 883–889.
doi:10.1016/j.jbiotec.2008.07.1203 VI6-P-021 Fractional factorial design and central composite design for CO2 mitigation using Senedesmus producto-capitatus Dong-Keon Kim ∗ , Ji-Sue Kwon, Seong-Joo Hong, Choul-Gyun Lee Institute of Industrial Biotechnology, Department of Biological Engineering, Inha University, Incheon 402-751, South Korea Carbon dioxide (CO2 ) is considered as a major greenhouse gas causing the global warming problem. Mankind has now recognized both the necessity and increasing urgency to reduce the CO2 content in the atmosphere. Carbon dioxide removal technology has two parts: biological CO2 fixation (BCF) and separation technology. BCF is environment-friendly way to remove CO2 and photosynthesis by microalgae and higher plants is the most common BCF. Since the growth rates of some microalgae are 30 times higher than higher plants, microalgae can be a feasible solution for biological CO2 mitigation. Several different microalgal strains have tested for the possible application in BCF. Among them, optimization of Scenedesmus producto-capitatus is reported here. The strain was cultured in different CO2 concentrations: air, 5% CO2 balanced with air, and 10% CO2 (+90% air). S. producto-capitatus grew well under higher CO2 concentration. Further statistical optimization on culture media using fractional factorial design (FFD) and central composite (CCD) performed for Scenedesmus in order to optimize biological CO2 fixation. From FFD, the four components, CaCl2 ·2H2 O, MgSO4 ·7H2 O, Fe-ammonium citrate and Na2 CO3 , were identified as major factors. In the subsequent experiment, a CCD was then applied with the four factors, at five levels each. The result showed the optimal condition as 0.15 g/L of MgSO4 ·7H2 O, and 0.08 g/L of CaCl2 ·2H2 O, 0.016 g/L of Fe-ammonium citrate and 0.36 g/L of Na2 CO3 . References Li, C., Bai, J., Cai, Z., Ouyang, F., 2002. Optimization of a cultural medium for bacteriocin production by Lactococcus lactis using response surface methodology. J. Biotechnol. 93 (January (1)), 27–34.
S596
Abstracts / Journal of Biotechnology 136S (2008) S589–S601
Purama, R.K., Goyal A., 2008. Screening and optimization of nutritional factors for higher dextransqucrase production by Leuconostomesenteroides NRRL B-640 using statistical approach. Bioresour. Technol.
doi:10.1016/j.jbiotec.2008.07.1204
VI6-P-0203 Extraction and properties of collagen from sea cucumber (Stichopus japonicus) body wall Xiuping Dong 1,2 , Beiwei Zhu 1,2,∗ , Yang Gao 1 , Da-yong Zhou 1 1
Free radical scavenging activity of gelatin hydrolysate from puffer fish skin, Takifugu rubripes
College of Bio & Food Technology, Dalian Polytechnic University, Dalian, 116034, PR China 2 School of Food, Biological Engineering, Jiangsu University, Zhenjiang, Jiangsu, 212013, China
Beiwei Zhu ∗ , Dayong Zhou, Dongmei Li, Junfeng Ren
E-mail address: [email protected] (B. Zhu).
College of Bio & Food Technology, Dalian Polytechnic University, Dalian, 116034, PR China
Sea cucumbers are economically important echinoderms that are widely cultured in China. The body wall is the main edible part of sea cucumber composed of epithelial tissue and dermal connective tissue. Sea cucumber body wall is capable of undergoing a high degree of autolysis in response to a variety of environmental and mechanical cues, such as UV exposure and lower salinity (Dai, 1990). There has been a rapid expansion and intensification of commercial sea cucumber farming in China, and the autolysis of sea cucumber has developed into a major problem in product processing and preservation. Though some investigators considered that this phenomenon was related to thermal degeneration of collagen in connective tissue, but they could not provide reliable evidence to prove it (Saito et al., 2002). Therefore, the isolation and characterization of gelatin from body wall of sea cucumber in combination with property studies may contribute to better understand the autolytic physiology of sea cucumbers. Image analysis of tissue slice using polarization microscope indicated that there are two type gelatins existing in connective tissue of sea cucumber. To determine the physicochemical characteristics of the two type gelatins, they were extracted from the body wall from sea cucumber. Gelatin I was pepsin soluble collagen. Taking yield rate as target, the optimum extraction condition was as below: temperature 4 ◦ C; pepsin addition 14% (w/w); dosage liquor ratio 1:500; extraction time 72 h. Under the optimum condition, the pepsin soluble collagen yield rate was 73.44%. FTIR spectroscopy confirmed that the pepsin soluble collagen have three-ply spiral structure. The weight average molecular weight of which was estimated to be 200–300 kDa by SDS-PAGE. According to the electrophoretic pattern, the collagen consisted of (a1 )2 a2 . Gelatin II was insoluble collagen, which was extracted according to the method of Frederick et al. (Thurmond and Trotter, 1996). Further characterization and property studies on the purified gelatin from this sea cucumber are currently in progress.
VI6-P-022
E-mail address: [email protected] (B. Zhu). Free radicals have been implicated in the etiology of many diseases, including inflammatory disease, cancer, diabetes, and aging (Pryor and Ann, 1982). In order to act against free radicals in food and biological system, many synthetic antioxidants have been commonly used. However, the applications of them are restricted due to potential hazards related to health (Grice, 1988). Therefore, enhancement of body’s antioxidant defenses through natural and safe antioxidants would seem to provide a reasonable and practical approach. Published data indicate that gelatins, the heterogeneous mixture of high molecular weight water-soluble protein derived from collagen and theirs hydrolysates in general have strong antioxidant activities. In present study, we investigate the radicals-scavenging activity of enzymatically prepared puffer fish skin collagen hydrolysates and isolated a potent radical-scavenging peptide. Puffer fish skin gelatin was hydrolyzed using papain and the resulting hydrolysate was fractionated through an ultrafiltration membrane system separately with three different molecular weight cut-offs (10, 3 and 1 kDa). The hydrolysate and its MW fractions (fraction 1: 3–10 kDa; fraction 2: 1–3 kDa; fraction 3: <1 kDa) were evaluated using hydroxyl radical scavenging assay and DPPH radical scavenging activity. The gelatin peptide from puffer fish skin exhibited hydroxyl radical and DPPH radical scavenging activities higher than that of Vc and Ve as positive control. Among hydrolysates, fraction 3 exhibited the highest antioxidant activities than those of other fractions. In order to purity a peptide having potent antioxidant properties, fraction 3 was separated using consecutive chromatographic methods on a DEAE Sepharose CL-6B anion exchange column, Sephadex G-25 gel filtration column and TSK-G4000PWXL gel column. Finally, a potent antioxidative peptide was purified to homogeneity with amino acid sequence was confirmed. References Grice, H.C., 1988. Safety evaluation of butylated hydroxyanisole from the perspective of effects on forestomach and oesophageal squamous epithelium. Food Chem. Toxicol. 26, 717–723. Pryor, W.A., Ann, N.Y., 1982. Free radical biology: xenobiotics, cancer, and aging. Acad. Sci. 393, 1–22.
doi:10.1016/j.jbiotec.2008.07.1205
References Dai, Z.Y., 1990. A review on autolysis in fish. J. Zhejiang Fish Coll. 9, 51–56. Thurmond, F., Trotter, J., 1996. Morphology and biomechanics of the microfibrillar network of sea cucumber dermis. J. Exp. Biol. 199 (Pt 8), 1817–1828. Saito, M., Kunisaki, N., Urano, N., Kimura, S., 2002. Collagen as the major edible component of seacucumber (Stichopus japonicus). J. Food Sci. 67, 1319–1322.
doi:10.1016/j.jbiotec.2008.07.1206
total carbohydrate content is, however, 10% lower than that of the control.
References Fidalgo, J.P., Cid, A., Torres, E., Sukenik, A., Herrero, C., 1998. Effects of nitrogen source and growth phase on proximate biochemical composition, lipid classes and fatty acid profile of the marine microalgae Isochrysis galbana. Aquaculture 166 (1998), 105–116. Jeffrey, S.W., Brown, M.R., Volkman, J.K., 1994. Haptophyte as feed stocks in mariculture. In: Green, J.C., Leadbeater, B.S.C. (Eds.), The Haptophyte Algae. Clarendon Press, Oxford, pp. 287–302.
doi:10.1016/j.jbiotec.2008.07.1201 VI6-P-017 A study on sponge associated actinomycetes and isolation of potent antimicrobial compounds V. Vaishnavapriya 1,∗ , R. Venkatesan 2 , N. Manoharan 3 1
Center for Ocean research, Sathyabama University, Chennai 119, India Ocean Science and Technology for Islands, National Institute of Ocean Technology, Chennai 601302, India 3 Sathyabama University, Chennai 119, India 2
E-mail address: [email protected] (V. Vaishnavapriya). Sponges are well known to harbor diverse microbes and represent a significant source of bioactive compounds. As far as the actinomycetes are concern, they are the prime producers of antibiotics. It is strongly believed that the microbes do produce the potent secondary metabolites at the extreme stress conditions which do possess potent antimicrobial activity. The increasing occurrence of infectious disease is becoming a world wide problem. Additionally, the resistance problem demands that the renewal efforts be made to seek antimicrobial agents that are effective against pathogenic microorganisms resistant to current treatment. So this work mainly aims to isolate sponge associated actinomycetes and is targeted to extract the potent antimicrobial activity against clinical strains. The sponge extract of 60 g/concentration is very effective against the pathogenic bacterium. The Streptomycetes sp. isolated from the sponge shows the inhibitory property against the clinical strains. doi:10.1016/j.jbiotec.2008.07.1202 VI6-P-018 Comparative study of N-trimethyl chitosan (TMC) and chitooligosaccharides on anti-radical scavenging activity A. Ozhan Aytekin ∗ , Shigeru Morimura Graduate School Science and Technology, Kumamoto University, 2-391 Kurokami, Kumamoto 860-8555, Japan E-mail address: [email protected] (A.O. Aytekin). Chitosan is a biopolymer and mainly found in chitin form in crustacean shells. Chitosan, which is copolymer consisting of beta-(1 → 4)-2-acetamido-d-glucose and beta-(1 → 4)-2-amino-dglucose, units, is linear and natural cationic polysaccharide. Although chitosan is biocompatible, biodegradable and non-toxic polymer, its solution has high viscosity and it is mainly dissolved in acidic solutions. Therefore, its derivatives and oligomers are considered recently (Kurita, 2006; Kim and Rajapakse, 2005). In this study, initially N-trimethyl chitosan (TMC) were produced from commercial chitosan (400 kDa) and chitosan oligomers (1 kDa). Then char-
S595
acterization of chitosan, chitosan oligomers and TMC were done by H NMR, HPLC and viscosimeter (Amidi et al., 2006). Antiradical scavenging activity on the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical of three different degree of quaternization (DQ) of TMC and two different chitosan depending on molecular weight were investigated (Lin and Chou, 2004). The results showed that, TMC has higher antiradical activity than chitosan and its oligomers. Lower DQ and molecular weight of TMC and chitosan have higher scavenging effect (SE), separately. According to positive control, which is ascorbic acid at 1 mg/ml, 95.5% SE is observed and at the same concentration TMC, chitosan (400 kDa) and chitosan oligomers (1 kDa) ranging between 75% and 85%; 10–15%; 27–35% SE, respectively. References Amidi, M., Romeijn, S.G., Borchard, G., Junginger, H.E., Hennink, W.E., Jiskoot, W., 2006. Preparation and characterization of protein-loaded N-trimethyl chitosan nanoparticles as nasal delivery system. J. Control. Release 111, 107–116. Kurita, K., 2006. Chitin and chitosan: functional biopolymers from marine crustaceans. Mar. Biotechnol. 1–24. Kim, S.K., Rajapakse, N., 2005. Enzymatic production and biological activities of chitosan oligosaccharides (COS): a review. Carbohyd. Polym. 62, 357–368. Lin, H.Y., Chou, C.C., 2004. Antioxidative activities of water-soluble disaccharide chitosan derivatives. Food Res. Int. 37 (9), 883–889.
doi:10.1016/j.jbiotec.2008.07.1203 VI6-P-021 Fractional factorial design and central composite design for CO2 mitigation using Senedesmus producto-capitatus Dong-Keon Kim ∗ , Ji-Sue Kwon, Seong-Joo Hong, Choul-Gyun Lee Institute of Industrial Biotechnology, Department of Biological Engineering, Inha University, Incheon 402-751, South Korea Carbon dioxide (CO2 ) is considered as a major greenhouse gas causing the global warming problem. Mankind has now recognized both the necessity and increasing urgency to reduce the CO2 content in the atmosphere. Carbon dioxide removal technology has two parts: biological CO2 fixation (BCF) and separation technology. BCF is environment-friendly way to remove CO2 and photosynthesis by microalgae and higher plants is the most common BCF. Since the growth rates of some microalgae are 30 times higher than higher plants, microalgae can be a feasible solution for biological CO2 mitigation. Several different microalgal strains have tested for the possible application in BCF. Among them, optimization of Scenedesmus producto-capitatus is reported here. The strain was cultured in different CO2 concentrations: air, 5% CO2 balanced with air, and 10% CO2 (+90% air). S. producto-capitatus grew well under higher CO2 concentration. Further statistical optimization on culture media using fractional factorial design (FFD) and central composite (CCD) performed for Scenedesmus in order to optimize biological CO2 fixation. From FFD, the four components, CaCl2 ·2H2 O, MgSO4 ·7H2 O, Fe-ammonium citrate and Na2 CO3 , were identified as major factors. In the subsequent experiment, a CCD was then applied with the four factors, at five levels each. The result showed the optimal condition as 0.15 g/L of MgSO4 ·7H2 O, and 0.08 g/L of CaCl2 ·2H2 O, 0.016 g/L of Fe-ammonium citrate and 0.36 g/L of Na2 CO3 . References Li, C., Bai, J., Cai, Z., Ouyang, F., 2002. Optimization of a cultural medium for bacteriocin production by Lactococcus lactis using response surface methodology. J. Biotechnol. 93 (January (1)), 27–34.
S596
Abstracts / Journal of Biotechnology 136S (2008) S589–S601
Purama, R.K., Goyal A., 2008. Screening and optimization of nutritional factors for higher dextransqucrase production by Leuconostomesenteroides NRRL B-640 using statistical approach. Bioresour. Technol.
doi:10.1016/j.jbiotec.2008.07.1204
VI6-P-0203 Extraction and properties of collagen from sea cucumber (Stichopus japonicus) body wall Xiuping Dong 1,2 , Beiwei Zhu 1,2,∗ , Yang Gao 1 , Da-yong Zhou 1 1
Free radical scavenging activity of gelatin hydrolysate from puffer fish skin, Takifugu rubripes
College of Bio & Food Technology, Dalian Polytechnic University, Dalian, 116034, PR China 2 School of Food, Biological Engineering, Jiangsu University, Zhenjiang, Jiangsu, 212013, China
Beiwei Zhu ∗ , Dayong Zhou, Dongmei Li, Junfeng Ren
E-mail address: [email protected] (B. Zhu).
College of Bio & Food Technology, Dalian Polytechnic University, Dalian, 116034, PR China
Sea cucumbers are economically important echinoderms that are widely cultured in China. The body wall is the main edible part of sea cucumber composed of epithelial tissue and dermal connective tissue. Sea cucumber body wall is capable of undergoing a high degree of autolysis in response to a variety of environmental and mechanical cues, such as UV exposure and lower salinity (Dai, 1990). There has been a rapid expansion and intensification of commercial sea cucumber farming in China, and the autolysis of sea cucumber has developed into a major problem in product processing and preservation. Though some investigators considered that this phenomenon was related to thermal degeneration of collagen in connective tissue, but they could not provide reliable evidence to prove it (Saito et al., 2002). Therefore, the isolation and characterization of gelatin from body wall of sea cucumber in combination with property studies may contribute to better understand the autolytic physiology of sea cucumbers. Image analysis of tissue slice using polarization microscope indicated that there are two type gelatins existing in connective tissue of sea cucumber. To determine the physicochemical characteristics of the two type gelatins, they were extracted from the body wall from sea cucumber. Gelatin I was pepsin soluble collagen. Taking yield rate as target, the optimum extraction condition was as below: temperature 4 ◦ C; pepsin addition 14% (w/w); dosage liquor ratio 1:500; extraction time 72 h. Under the optimum condition, the pepsin soluble collagen yield rate was 73.44%. FTIR spectroscopy confirmed that the pepsin soluble collagen have three-ply spiral structure. The weight average molecular weight of which was estimated to be 200–300 kDa by SDS-PAGE. According to the electrophoretic pattern, the collagen consisted of (a1 )2 a2 . Gelatin II was insoluble collagen, which was extracted according to the method of Frederick et al. (Thurmond and Trotter, 1996). Further characterization and property studies on the purified gelatin from this sea cucumber are currently in progress.
VI6-P-022
E-mail address: [email protected] (B. Zhu). Free radicals have been implicated in the etiology of many diseases, including inflammatory disease, cancer, diabetes, and aging (Pryor and Ann, 1982). In order to act against free radicals in food and biological system, many synthetic antioxidants have been commonly used. However, the applications of them are restricted due to potential hazards related to health (Grice, 1988). Therefore, enhancement of body’s antioxidant defenses through natural and safe antioxidants would seem to provide a reasonable and practical approach. Published data indicate that gelatins, the heterogeneous mixture of high molecular weight water-soluble protein derived from collagen and theirs hydrolysates in general have strong antioxidant activities. In present study, we investigate the radicals-scavenging activity of enzymatically prepared puffer fish skin collagen hydrolysates and isolated a potent radical-scavenging peptide. Puffer fish skin gelatin was hydrolyzed using papain and the resulting hydrolysate was fractionated through an ultrafiltration membrane system separately with three different molecular weight cut-offs (10, 3 and 1 kDa). The hydrolysate and its MW fractions (fraction 1: 3–10 kDa; fraction 2: 1–3 kDa; fraction 3: <1 kDa) were evaluated using hydroxyl radical scavenging assay and DPPH radical scavenging activity. The gelatin peptide from puffer fish skin exhibited hydroxyl radical and DPPH radical scavenging activities higher than that of Vc and Ve as positive control. Among hydrolysates, fraction 3 exhibited the highest antioxidant activities than those of other fractions. In order to purity a peptide having potent antioxidant properties, fraction 3 was separated using consecutive chromatographic methods on a DEAE Sepharose CL-6B anion exchange column, Sephadex G-25 gel filtration column and TSK-G4000PWXL gel column. Finally, a potent antioxidative peptide was purified to homogeneity with amino acid sequence was confirmed. References Grice, H.C., 1988. Safety evaluation of butylated hydroxyanisole from the perspective of effects on forestomach and oesophageal squamous epithelium. Food Chem. Toxicol. 26, 717–723. Pryor, W.A., Ann, N.Y., 1982. Free radical biology: xenobiotics, cancer, and aging. Acad. Sci. 393, 1–22.
doi:10.1016/j.jbiotec.2008.07.1205
References Dai, Z.Y., 1990. A review on autolysis in fish. J. Zhejiang Fish Coll. 9, 51–56. Thurmond, F., Trotter, J., 1996. Morphology and biomechanics of the microfibrillar network of sea cucumber dermis. J. Exp. Biol. 199 (Pt 8), 1817–1828. Saito, M., Kunisaki, N., Urano, N., Kimura, S., 2002. Collagen as the major edible component of seacucumber (Stichopus japonicus). J. Food Sci. 67, 1319–1322.
doi:10.1016/j.jbiotec.2008.07.1206