[논문]솔잎 퇴적물에서 추출한 방향족 탄화수소물질 분해 박테리아의 동정

송윤재

솔잎 퇴적물에서 추출한 방향족 탄화수소물질 분해 박테리아의 동정
Characterization of Aromatic Hydrocarbon Degrading Bacteria Isolated from Pine Litter 원문보기

한국미생물·생명공학회지 = Korean journal of microbiology and biotechnology, v.37 no.4, 2009년, pp.333 - 339

송윤재 (경원대학교 생명과학과)

초록
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새로운 pine needle agar를 이용하여 15종의 박테리아를 솔잎퇴적물에서 추출하여 동정하였다. 이들 박테리아는 lignin biodegradation에서 주로 유도되는 방향족 탄화수소물질을 $\beta$-ketoadipate pathway의 ortho-cleavage를 이용하여 분해하는 것으로 밝혀졌다. 나아가서 이들 박테리아에 의한 여러 종의 방향족 탄화수소물질 분해에 관해서도 조사하였다. 본 연구는 솔잎 퇴적물에 존재하는 박테리아 종들이 방향족 탄화수소물질을 분해할 수 있는 대사능력을 가지고 있다는 것을 검증하였다.

Abstract ▼ AI-Helper

Using a novel pine needle agar, fifteen bacterial species were isolated from pine litter. These bacteria were able to degrade aromatic hydrocarbons derived from lignin and utilize the ortho-cleavage of the $\beta$-ketoadipate pathway to degrade protocatechuate or catechol. A different utilization array of aromatic hydrocarbons by these bacteria was also determined. This study provides the information on bacterial species living in pine litter and suggests that these bacteria have metabolic abilities to utilize aromatic hydrocarbons derived from lignin biodegradation.

주제어

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문제 정의

The purpose of this research was to isolate bacteria from pine litter and to examine the evidence for their involvement in lignin biodegradation. Pine Needle Agar (PNA) was used as a selective medium to isolate bacteria from pine litter.

제안 방법

Bacteria living in pine litter may be involved in lignin recycling by utilizing low molecular weight aromatic hydrocarbons derived from fungal digestion of lignin. In this study, bacterial flora from pine litter were isolated and characterized for their metabolic capacities to utilize aromatic monomers derived from lignin biodegradation.

대상 데이터

Since a variety of aromatic hydrocarbons are degraded by catabolic pathways that converge upon protocatechuate or catechol under aerobic conditions, the ability of these isolates to grow with protocatechuate or catechol was determined. Among the forty three isolates initially obtained, fifteen isolates were able to utilize both protocatechuate and catechol as an energy source or a carbon source. Doubling times ranged from 2.
To determine whether the bacterial isolates from pine litter have metabolic capacities to utilize aromatic hydrocarbons, the growth of the isolates with low molecular weight aromatic hydrocarbons derived from lignin was tested (Table 2). The aromatic hydrocarbons tested in this study were: protocatechuate, catechol, hydroquinone, phloroglucinol, phenyl acetate, salicylate, gentisate, pyrogallol, paminobenzoic acid, vanillate, gallic acid, resorcinol, p hydroxybenzoic acid and phenyl benzoate [15]. Table 2 lists the result of growth tests of the isolates with these aromatic hydrocarbons.
After 2-3days of incubation (Klett units between 100-150), the cells were transferred into 18mm diameter tubes containing DMM with 17 mM of a selected low molecular weight aromatic hydrocarbons. The aromatic hydrocarbons used for this test were: protocatechuate, catechol, pyrogallol, resorcinol, gallic acid, hydroquinone, p-aminobenzoic acid, salicylate, gentisic acid, vanillate, p-hydroxybenzoic acid, and phenyl acetate. After 10-15 days of incubation, visible turbidity was taken as evidence of growth.
Pine litter samples were collected at Hartmann’s Creek State Forest, west of Waupaca, Wisconsin. The collection site was about 1.5 km into the forest and about fifty meters from a nature trail. Sterile forceps were used to place 5.

이론/모형

A reference cuvette contained all components except substrate. Concentration of absorbing material in the sample was calculated by using the BeerLambert law. The coefficient was obtained by preparing a standard curve and was found to be 4,230.
To determine whether the bacterial isolates from pine litter degrade protocatechuate or catechol by the ortho- or meta-cleavage, the Rothera test was performed as described in materials and methods. All isolates that were able to utilize either protocatechuate or catechol converted substrates to β-ketoadipate by the ortho-cleavage (data not shown).

성능/효과

2). Pseudomonas fluorescens species had higher levels of protocatechuate3,4-oxygenase activities than other isolates, and Microbacterium esteraromaticum had a higher level of catechol1,2-oxygenase activity than other isolates.
A BIOLOG analysis was also used as an independent method to identify the isolates. The bacterial isolates were identified as Pseudomonas marginalis, Rhodococcus fascians, Pseudomonas fragi, Pseudomonas corrugata, Serratia marcescens, Flavimonas oryzihabitans, Klebsiella pneumoniae ozaenae, Pseudomonas putida type B1, Pantoea dispersa, Burkholderia cepacia, Pseudomonas fluorescens Type B, F and G, Serratia liquefaciens, and Microbacterium esteraromaticum. Most of these isolates were members of the genera Pseudomonas.

참고문헌 (25)

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