臺灣北部沿岸海水中甲醛降解微生物的分離和特性研究

Isolation and characterization of formaldehyde-degrading microorganisms in coastal seawater of north Taiwan

周振鎏(Chan-Lao Chau)

指導教授 ： 侯文祥

共同指導教授 ： 謝文陽(Wung-Yang Shieh)

國立臺灣大學/生物資源暨農學院/生物環境系統工程學研究所/碩士(2013年)

https://doi.org/10.6342/NTU.2013.00205

摘要

甲醛 ( formaldehyde ) 是一種廣泛應用於醫療、農業、工業的化學物，因其可破壞蛋白質和核酸，故具有生物毒性。本研究於台灣北部沿岸 5 個地點採集海水樣本，進行甲醛耐受微生物的培養、計數和分離，並測試分離株是否具甲醛降解能力以及研究其降解效率。結果發現，各樣水中甲醛耐受微生物的菌數均大於 240 cells / ml。根據 16S rRNA gene 及 26S rRNA gene 的定序及比對，顯示 8 株可耐受 25 ppm 甲醛的分離株中，4 株為真菌，分別為 Galactomyces 屬（1 株）及 Candida 屬（3 株），另外 4 株為細菌，分別為 Enterobacter 屬（1 株）、 Pseudomonas 屬（1 株）及 Bacillus 屬（2 株）。所有菌株皆無法以甲醇作為單一碳源生長。除 2 株 Bacillus 屬細菌，其餘 6 株分離株具有較強的甲醛耐受能力，以之進行甲醛降解測試，當菌液起始濃度為 0.2 ( OD600 ) 時，可在 PY broth 中於 72 小時內分解 54 % 至 98 % 100 ppm 甲醛，而細菌分離株 B3 在 PY broth 中可於 48小時內分解 100 % 400 ppm 甲醛，於 72 小時內分解 59 % 700 ppm 甲醛，提升 B3 的菌液起始濃度成 0.55 至 0.6 ( OD600 ) 並以搖瓶方式培養，其於 24 小時內即可分解約 97 % 1500 ppm 或 60 % 3200 ppm 甲醛。綜合分子親緣分析及生理生化特性測試結果，推斷 B3 屬 Enterobacter cloacae。分離株 B3 於未來或可應用於處理甲醛廢水。

關鍵字

甲醛 ； 生物降解 ； 甲基營養型 ； 腸桿菌屬 ； 陰溝腸桿菌

並列摘要

Formaldehyde is a widely used chemical in medicine, agriculture and different industrial areas. It is biologically toxic because of its proteins and nucleic acid denaturing properties.
Coastal seawater samples were collected from 5 regions of northern Taiwan. Microorganisms were cultured, counted and isolated for studies of formaldehyde degradation ability and degradation efficiency. Eight formaldehyde-tolerant bacterial and fungal strains were isolated. All strains can tolerate up to 25 ppm formaldehyde, but cannot grow on methanol as a sole carbon source. The formaldehyde-tolerant microorganisms were found in all water samples with concentrations greater than 240 cells / ml. The fungal isolates were classified to the genus Galactomyces ( 1 strain ) or Candida ( 3 strains ).The bacterial isolates were classified to the genus Enterobacter ( 1 strain ), Pseudomonas ( 1 strain ) or Bacillus ( 2 strains ). Isolates ( except 2 strains of Bacillus ) were used for formaldehyde biodegrading study, 54 % ~ 98 % of 100 ppm formaldehyde were degraded by these strains over 72 hours when the culture’s initial optical density at 600 nm was 0.2. 100 % of 400 ppm or 59 % of 700 ppm formaldehyde in PY broth were degraded by strain B3 in 48 or 72 hours. Strain B3 could also degrade 97 % of 1500 ppm or 60 % 3200 ppm formaldehyde in PY broth over 24 hours when the culture’s initial optical density at 600 nm was 0.55 ~ 0.6, and incubated with shaking. 16S rRNA gene sequence and phenotype analysis indicated that strain B3 was Enterobacter cloacae. Strains B3 may be useful for treatment of waste water containing formaldehyde.

並列關鍵字

Formaldehyde ； biodegradation ； methylotrophic ； Enterobacter cloacae

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