ISOLATION AND OPTIMIZATION OF PAH-DEGRADATIVE BACTERIA

FROM CONTAMINATED SOIL FOR PAHs BIOREMEDIATION

J. W. C. WONG∗ , K. M. LAI, C. K. WAN, K. K. MA and M. FANG

Department of Biology, Hong Kong Baptist University, Kowloon Tong, Hong Kong
(∗ author for correspondence, e-mail: jwcwong@hkbu.edu.hk, fax: 852 2339 5995)

(Received 20 March 2000; accepted 9 October 2001)

Abstract. The objective of the present study is to isolate PAH-degradative bacteria from petroleum-
contaminated soils and to optimize their degradative conditions including pH, glucose, nitrogen and
phenanthrene concentrations required for bioremediation purposes. Several bacterial strains were
isolated through enrichment and one strain, Burkholderia cocovenenans (BU-3) that was tentatively
identified by the Biolog system, demonstrated a high removal rate of phenanthrene over other strains.
More than 95% in 100 and 500 mg L−1 and 65% in 1000 mg L−1 of phenanthrene contents was
reduced in the culture media, respectively. Maximum rate of phenanthrene removal up to 4.2 mg
hr−1 occurred in the culture containing 1000 mg L−1 phenanthrene. Media at a pH between 6.5
to 7.0 were more favorable for the degradation of phenanthrene by BU-3. Although increasing
glucose concentrations from 0.45 to 3 g L−1 resulted in a better bacterial growth of the isolated
bacteria, the degradation of phenanthrene was reduced significantly. Nitrogen supplement did not
exert a significant effect on bacterial growth and phenanthrene degradation. The isolated Burkhold-
eria cocovenenans BU-3 demonstrated to be a feasible strain for degradation of phenanthrene at a
neutral pH, even up to a phenanthrene concentration of 1000 mg L−1 .

Keywords: bioremediation, Burkholderia cocovenenans, degradation, glucose, pH, phenanthrene

1. Introduction

Remediation of petroleum-contaminated soils has received an increasing concern

in China, because large areas of petroleum-contaminated soil are posing threats to
the ecosystem and human health. A major fraction of petroleum mixtures, poly-
cyclic aromatic hydrocarbons (PAHs), are recalcitrant to microbial attack and have
highly toxic, mutagenic or carcinogenic effects. Hence, PAHs in contaminated soils
must be treated to avoid any possible hazard to human health. Chemical, physical
and biological methods have been utilized to remediate PAH-contaminated soils.
However, the principal processes for the removal of PAHs from the environment are
thought to be microbial transformation and degradation. Bioremediation, which is
based on certain species of microorganisms to metabolize PAHs either completely
or partially, is a cheap and effective means to decontaminate PAH-contaminated
soils (Hinchee et al., 1994).
Many heterogeneous or heterotrophic microorganisms, found in aqueous envir-
onments, soils or sediments, can use PAH compounds as a sole source of carbon

Water, Air, and Soil Pollution 139: 1–13, 2002.

© 2002 Kluwer Academic Publishers. Printed in the Netherlands.