Production of Medium-chain-length Poly (3-hydroxyalkanoates) by Pseudomonas sp. EML8 from Waste Frying Oil

Taegyeong Kim, Jong-Sik Kim, Chung-Wook Chung
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Abstract

In this study, to reduce the production cost of poly(3-hydroxyalkanoates) (PHA), optimal cell growth and PHA biosynthesis conditions of the isolated strain Pseudomonas sp. EML8 were established using waste frying oil (WFO) as the cheap carbon source. Gas chromatography (GC) and GC mass spectrometry analysis of the medium-chain-length PHA (mcl-PHAWFO) obtained by Pseudomonas sp. EML8 of WFO indicated that it was composed of 7.28 mol% 3-hydrxoyhexanoate, 39.04 mol% 3-hydroxyoctanoate, 37.11 mol% 3-hydroxydecanoate, and 16.58 mol% 3-hydroxvdodecanoate monomers. When Pseudomonas sp. EML8 were culture in flask, the maximum dry cell weight (DCW) and the mclPHAWFO yield (g/l) were showed under WFO (20 g/l), (NH4)2SO4 (0.5 g/l), pH 7, and 25°C culture conditions. Based on this, the highest DCW, mcl-PHAWFO content, and mcl-PHAWFO yield from 3-l-jar fermentation was obtained after 48 hr. Similar results were obtained using 20 g/l of fresh frying oil (FFO) as a control carbon source. In this case, the DCW, the mcl-PHAFFO content, and the mcl-PHAFFO yields were 2.7 g/l, 62 wt%, and 1.6 g/l, respectively. Gel permeation chromatography analysis confirmed the average molecular weight of the mcl-PHAWFO and mcl-PHAFFO to be between 165-175 kDa. Thermogravimetric analysis showed decomposition temperature values of 260°C and 274.7°C for mclPHAWFO and mcl-PHAFFO, respectively. In conclusion, Pseudomonas sp. EML8 and WFO could be suggested as a new candidate and substrate for the industrial production of PHA.
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假单胞菌sp. EML8利用废煎炸油制备中链长聚3-羟基烷酸酯
为了降低聚(3-羟基烷酸酯)(PHA)的生产成本,本研究以废煎炸油(WFO)为廉价碳源,建立了分离菌株Pseudomonas sp. EML8的最佳细胞生长和PHA生物合成条件。由假单胞菌sp. EML8从WFO中分离得到的中链PHA (mcl- phofo)经气相色谱和质谱分析表明,其主要由7.28 mol% 3-羟基己酸酯、39.04 mol% 3-羟基辛酸酯、37.11 mol% 3-羟基癸酸酯和16.58 mol% 3-羟基十二酸酯单体组成。假单胞菌EML8在烧瓶中培养时,在WFO (20 g/l)、(NH4)2SO4 (0.5 g/l)、pH 7和25℃的培养条件下,获得了最大干细胞重(DCW)和mclPHAWFO产量(g/l)。在此基础上,3-l-罐发酵48h后,DCW、mcl-PHAWFO含量和mcl-PHAWFO产量最高。用20 g/l的新鲜煎炸油(FFO)作为对照碳源,得到了类似的结果。在这种情况下,DCW、mcl-PHAFFO含量和mcl-PHAFFO产率分别为2.7 g/l、62 wt%和1.6 g/l。凝胶渗透色谱分析证实mcl-PHAFFO和mcl-PHAFFO的平均分子量在165-175 kDa之间。热重分析表明,mclphaffo和mcll - phaffo的分解温度分别为260℃和274.7℃。综上所述,假单胞菌sp. EML8和WFO可作为PHA工业生产的新候选物和底物。
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