• Journal of Microbiology
• /
• v.41 no.4
• /
• pp.349-352
• /
• 2003

Recent studies have shown that some of the PCB (polychlorinated biphenyl)-degraders are able to effectively degrade PCB in the presence of monoterpenes, which act as inducers for the degradation pathway. Rhodococcus sp. T104, an effective PCB degrader, has been shown to induce the degradation pathway by utilizing limonenes, cymenes, carvones, and pinenes as sole carbon sources which can be found in the natural environment. Moreover, the strain T104 proved to possess three separate oxidation pathways of limonene, biphenyl, and phenol. Of these three, the limonene can also induce the biphenyl degradation pathway. In this work, we report the presence of three distinct genes for aromatic oxygenase, which are putatively involved in the degradation of aromatic substrates including biphenyl, limonene, and phenol, through PCR amplification and denaturing gradient gel electrophoresis (DGGE). The genes were differentially expressed and well induced by limonene, cymene, and plant extract A compared to biphenyl and/or glucose. This indicates that substrate specificity must be taken into account when biodegradation of the target compounds are facilitated by the plant natural substrates.

• Korean Journal of Environmental Agriculture
• /
• v.42 no.4
• /
• pp.259-268
• /
• 2023

Rhodococcus sp. 3-2 strain has been reported to degrade benzimidazole-based pesticides, such as benomyl and carbendazim. Therefore, this study aimed to optimize culture medium composition and culture conditions to achieve cost-effective and efficient large-scale production of the Rhodococcus sp. 3-2 strain. The study identified that the optimal media composition for mass culture comprised 0.5% glucose, 0.5% yeast extract, 0.15% NaCl, 0.5% K₂HPO₄, 0.5% sodium succinate, and 0.1% MgSO₄. Additionally, a microbial agent was developed using a 1.5-ton fermenter, with skim milk (20%), monosodium glutamate (15%), and vitamin C (2%) as key components. The storage stability of the microbial agent has been confirmed, with advantages of low temperature conservation, which helps to sustain efficacy for at least six months. We also assessed the benomyl degradation activity of the microbial agent within field soil. The results revealed an over 90% degradation rate when the concentration of viable cells exceeded 2.65 × 10⁶ CFU/g after a minimum of five weeks had elapsed. Based on these findings, Rhodococcus sp. 3-2 strain can be considered a cost-effective microbial agent with diverse agricultural applications.

• Journal of Environmental Science International
• /
• v.11 no.6
• /
• pp.553-560
• /
• 2002

The research was performed to compare to the biofilm characteristics and phenol removal efficiency in RBCs(Rotating Biological Contactor) using Rhodococcus sp. EL-GT(single population) and activated sludge(mixed population) as inoculum. Both reactors showed similar tendency on variations of dry weight, thickness and dry density of biofilm. However, the growth of biofilm thickness in 3 and 4 stage of single population reactor has sustained longer than that of the mixed population reactor. Unlike the mixed population reactor, the dry density of biofilm in the single population reactor had a difference between 1, 2 stage and 3, 4 stage. The single population reactor was stably operated without the decrease of phenol removal efficiency in the range of pH 6 ~ 9 and 15mM phenol was completely degraded in these pH ranges. But in case of the mixed population reactor, the phenol degradability was dramatically decreased at over 5mM phenol concentration because of the overgrowth and detachment of its biofilm.

• Proceedings of the Korean Environmental Sciences Society Conference
• /
• 2001.11a
• /
• pp.102-104
• /
• 2001

본 연구는 방향족 화합물질 중 페놀폐수에 대한 생물학적 처리를 위해 본 실험실에서 분리한 페놀분해능이 우수한 Rhodococcus sp. EL-GT를 이용하여 catechol 분해 catechol 1,2-dioxygenase 분해활성을 측정하였고, 이것이 ortho-pathway임을 확인할 수 있었다. 또한 다른 연구에서 보고된 Rhodococrus rhodochrous NCIMB 13259 균주의 catechol 1,2 dioxygenase를 기초로한 primer를 이용하여 PCR을 수행하였으며 이 분해 유전자의 cloning실험을 수행 중이다. 이들 실험을 통하여 Rhodoroccus sp. EL-GT의 페놀분해 균의 유전적 구조 및 특성을 검토하고 이를 이용하여 방향족 화합물의 분해능이 보다 우수한 균주의 개발을 시도하고자 한다.

• Proceedings of the Microbiological Society of Korea Conference
• /
• 2008.05a
• /
• pp.164-164
• /
• 2008

• Journal of Microbiology and Biotechnology
• /
• v.30 no.6
• /
• pp.937-945
• /
• 2020

N-acyl-homoserine lactone (AHL)-mediated quorum sensing (QS) plays a major role in development of biofilms, which contribute to rise in infections and biofouling in water-related industries. Interference in QS, called quorum quenching (QQ), has recieved a lot of attention in recent years. Rhodococcus spp. are known to have prominent quorum quenching activity and in previous reports it was suggested that this genus possesses multiple QQ enzymes, but only one gene, qsdA, which encodes an AHL-lactonase belonging to phosphotriesterase family, has been identified. Therefore, we conducted a whole genome sequencing and analysis of Rhodococcus sp. BH4 isolated from a wastewater treatment plant. The sequencing revealed another gene encoding a QQ enzyme (named jydB) that exhibited a high AHL degrading activity. This QQ enzyme had a 46% amino acid sequence similarity with the AHL-lactonase (AidH) of Ochrobactrum sp. T63. HPLC analysis and AHL restoration experiments by acidification revealed that the jydB gene encodes an AHL-lactonase which shares the known characteristics of the α/β hydrolase family. Purified recombinant JydB demonstrated a high hydrolytic activity against various AHLs. Kinetic analysis of JydB revealed a high catalytic efficiency (k_cat/K_M) against C4-HSL and 3-oxo-C6 HSL, ranging from 1.88 x 10⁶ to 1.45 x 10⁶ M^-1 s^-1, with distinctly low K_M values (0.16-0.24 mM). This study affirms that the AHL degrading activity and biofilm inhibition ability of Rhodococcus sp. BH4 may be due to the presence of multiple quorum quenching enzymes, including two types of AHL-lactonases, in addition to AHL-acylase and oxidoreductase, for which the genes have yet to be described.

• Microbiology and Biotechnology Letters
• /
• v.37 no.3
• /
• pp.197-203
• /
• 2009

An indole oxygenase originated from Rhodococcus sp. RHA1 was cloned into the expression vector, pTrc99A, in Escherichia coli, and designated pTCAN1. The pTCAN2 was constructed from pTCAN1 by the deletion of $lacI^q$ for the constitutive expression of indole oxygenase without adding IPTG in the medium. The complete open reading frame of indole oxygenase was 1,224 bp long, which encodes a protein of 407amino acids. Crude extracts of E. coli $DH5{\alpha}$/pTCAN1 and pTCAN2, respectively, were prepared and subjected to SDS-PAGE analysis. A band corresponding to molecular mass of about 43 kDa was appeared and this result correlated with the predicted molecular mass of cloned indole oxygenase. The E. coli harboring pTCAN1 and pTCAN2, respectively, showed blue color colony in LB plate. The pigment showing blue color was prepared from E. coli $DH5{\alpha}$/pTCAN2, and identified as indigo by experiments using spectrophotometer, HPLC, and TLC. The indigo-forming activity of indole oxygenases from the whole cell of E. coli $DH5{\alpha}/pTCAN1$ cultured at LB medium added 1mM of IPTG and that of E. coli/pTCAN2 showed about 1.75nmol/min/mg DCW (dry cell weight) and 3.85 nmol/min/mg DCW, respectively. Also, the E. coli $DH5{\alpha}$/pTCAN2 produced about $236{\mu}M$ of indigo after 48 hours incubation in TB medium supplemented with 2.5 mM of tryptophan.

• Korean Journal of Microbiology
• /
• v.36 no.4
• /
• pp.279-284
• /
• 2000

Simultaneous reduction of Cr(VI) and degradation of phenol was observed in batch and bench-scale continuous stirred tank reactors using Rhodococcus sp. CP01 isolated from leachate. The strain CP01, which was capable of utilizing phenol as a sole source of carbon and energy, completely reduced added hexavalent chromium (0.25 mM) to its trivalent form during 60 hr batch assay under optimal conditions (pH 7.0 and 1,000 mg/L of phenol concentration). The rates of Cr(VI) reduction and phenol degradation were estimated as 4.17 $\mu$M Cr(VI) and 38.4 mg phenol.$L^{-1}{\cdot}hr^{-1}$, respectively. The continuous culture experiment was conducted for 46 days using synthetic feed containing different levels of chromate (0.0625 to 0.25 mM) and phenol(1,000 to 4,000 mg/L). With a hydraulic retention time of 100 hr, Cr(VI) reduction efficiency was mostly 100% for influent Cr(VI) and phenol concentrations of 0.125 mM and 3,000 mg/L, respectively. During quasi-steady-state operation, specific rate of Cr(VI) reduction was calculated as 0.34 mg Cr(VI).g $protein^{-1}{\cdot}hr^{-1}$ which was comparable to reported values obtained by using glucose as growth substrate. The results suggest the potential application of biological treatment for detoxification of wastewater contaminated simultaneously with Cr(VI) and pheonol.

• Korean Journal of Microbiology
• /
• v.49 no.3
• /
• pp.297-300
• /
• 2013

EPS producing bacteria were enumerated in ginseng root system (rhizosphere soil, rhizoplane, inside of root). EPS producing bacterial density of rhizosphere soil, rhizoplane and inside of root were distributed $9.0{\times}10^6$ CFU/g, $7.0{\times}10^6$ CFU/g, and $1.4{\times}10^3$ CFU/g, respectively. Phylogenetic analysis of the 24 EPS producing isolates based on the 16S rRNA gene sequences, EPS producing isolates from rhizosphere soil (RS) belong to genus Arthrobacter (6 strains) and Rhizobium (1 strain). EPS producing bacteria from rhizoplane (RP) were Arthrobacter (6 strains), Rhodococcus (1 strain) and Pseudomonas (1 strain). EPS producing bacteria from inside of root (IR) were categorized into Rhzobium (6 strains), Bacillus (1 strain), Rhodococcus (1 strain), and Pseudomonas (1 strain). Phylogenetic analysis indicated that Arthrobacter may be a member of representative EPS producing bacteria from ginseng rhizosphere soil and rhizoplane, and Rhizobium is typical EPS producing isolates from inside of ginseng root. The yield of EPS was 10.0 and 4.9 g/L by Rhizobium sp. 1NP2 (KACC 17637) and Arthrobacter sp. 5MP1 (KACC 17636). The purified EPS were analyzed by Bio-LC and glucose, galactose, mannose and glucosamine were detected. The major EPS sugar of these strains was glucose (72.7-84.9%).

• Journal of Microbiology and Biotechnology
• /
• v.23 no.2
• /
• pp.260-266
• /
• 2013

Rhodococcus sp. strain p52, a previously isolated dibenzofuran degrader, could effectively inhibit the growth of cyanobacteria, including species of Microcystis, Anabaena, and Nodularia. When strain p52 was inoculated at the concentration of $7.7{\times}10^7$ CFU/ml, 93.5% of exponentially growing Microcystis aeruginosa ($7.3{\times}10^6$ cells/ml initially) was inhibited after 4 day. The threshold concentration for its algicidal activity against M. aeruginosa was $7.7{\times}10^6$ CFU/ml. Strain p52 exerted algicidal effect by synthesizing extracellular substances, which were identified as trans-3-indoleacrylic acid, DL-pipecolic acid, and L-pyroglutamic acid. The effective concentrations of trans-3-indoleacrylic acid and DL-pipecolic acid against M. aeruginosa were tested to be 0.5 mg/l and 5 mg/l, respectively.