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http://ir.cmu.edu.tw/ir/handle/310903500/28923
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| 題名: | Production of recombinant porcine lactoferrin exhibiting antibacterial activity in methylotrophic yeast, Pichia pastoris |
| 作者: | Chen, HL;Lai, YW;Yen, CC;Lin, YY;Lu, CY;Yang, SH;Tsai, TC;Lin, YJ;Lin, CW;Chen, CM |
| 貢獻者: | 北港附設醫院;Natl Chung Hsing Univ, Dept Life Sci, Taichung 402, Taiwan;Da Yeh Univ, Dept Mol Biotechnol, Changhua, Taiwan;Natl Chung Hsing Univ, Ctr Biotechnol, Taichung 402, Taiwan;China Med Univ Peikang Hosp, Peikang, Taiwan;Chung Shang Med Univ, Dept Dent, Taichung, Taiwan;Natl Taiwan Univ, Dept Anim Sci, Taipei 10764, Taiwan |
| 關鍵詞: | Pichia pastoris;Recombinant lactoferrin |
| 日期: | 2004 |
| 上傳時間: | 2010-09-24 14:14:06 (UTC+8) |
| 出版者: | KARGER |
| 摘要: | Background Obesity is a risk factor for multiple chronic diseases. Aim To determine the relationship between overall or abdominal obesity and various metabolic abnormalities. Methods The medical records of participants undergoing examination from January 2000 to December 2002 were analysed. Body mass index (BMI; kg/m(2)) for overall obesity and waist circumference (WC; cm) indicating abdominal obesity were measured. Four groups were studied: (1) BMI < 27kg/m(2), WC: female < 80cm or male < 90cm; (2) BMI? 27kg/m(2), WC: female < 80cm or male < 90cm; (3) BMI < 27kg/m(2), WC: female greater than or equal to80cm or male greater than or equal to90cm; (4) BMI greater than or equal to 27kg/m(2), WC: female greater than or equal to 80cm or male greater than or equal to 90cm. Results There were 1,342 (44%) females and 1,711 males aged 20-87 years. The prevalence of overall obesity was 20.4% and abdominal obesity was 48%. Obese subjects had more metabolic abnormalities than non-obese in terms of risk of hypertension, hyperglycaemia, hypercholesterolaemia, hypertriglyceridaemia, high level of LDL, low level of HDL, high ratio of TC/HDL, hyperuricaemia and fatty liver. Conclusions As overall and abdominal obesity may predict clustering of metabolic abnormalities we suggest that BMI and WC are convenient methods for assessing metabolic abnormalities in primary healthcare settings.
Lactoferrin is a metal-binding glycoprotein exhibiting multifunctional immunoregulation of antibacterial, antioxidant, anti-endotoxin and antiviral activities. Uptake of porcine lactoferrin (PLF) has been shown to enhance resistance to diarrhea and anemia in neonatal piglets. In this study, the methylotrophic yeast, Pichia pastoris, was used to express a recombinant PLF (rPLF) gene from swine mammary gland. A synthetic secretion cassette was constructed using the inducible promoter of the alcohol oxidase-1 gene (AOX1) and the yeast α-mating factor signal peptide. After electroporation and Zeocin selection, several clones expressed high levels of rPLF protein which constitutes more than 30% of the total protein. A time-course study showed that rPLF mRNA transcripts are stably expressed during 120 h of culture induction. rPLF was exported into the culture supernatant at approximately 87 mg/l and a large portion of rPLF was accumulated in the cell cytoplasm at approximately 760 mg/l after 72 h of methanol induction. Recombinant PLF protein was purified via a heparin column using a fast protein liquid chromatography system. The glycosylation of P. pastoris-derived rPLF was analyzed and similar patterns to milk PLF were observed. Pepsin hydrolysate of rPLF displayed high bactericidal activity against Escherichia coli ATCC 25922 under scanning electron microscopy observation and minimal inhibitory concentration and minimal bactericidal concentration tests. Our results suggested that the methylotrophic yeast-inducible system is suitable for large-scale production of active antibacterial rPLF glycoprotein. |
| 關聯: | JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY 8(3):141-149 |
| 顯示於類別: | [北港附設醫院] 期刊論文
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