• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.541-544
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• 2001

One of the most promising TCE treatment systems is trickling biofilter (TBF), in which monooxygenase, the corresponding enzyme for initiating growth substrate oxidation, fortuitously transforms TCE via cometabolism. TCE. however. is not easily treated by simple cometabolic biotransformation. This is mainly due to the toxicity of TCE to microbial cell and monooxygenase. In this study, we cleveloped and operated a two-stage CSTR/TBF system for the long-term continuous treatment of TCE. In the two-stage biotransfon11ation system, CSTR with cell recycle from TBR was coupled to the TBR for the reactivation of the cells deactivated during TCE degradation.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.271-279
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• 2015

Lactoferrin (LF) is a multifunctional, iron-binding glycoprotein found in physiological secretions of mammals. LF shows antibacterial, antiviral and antifungal activities. In the present study, a gene encoding the N-terminal lobe of human lactoferrin (hLF) was isolated, cloned and expressed in methylotrophic yeast, Pichia pastoris. The recombinant hLF-N (rhLF-N) protein was secreted into the culture medium at the level of $458{\mu}g/ml$ in 3 L fermentor. The size of purified hLF-N was estimated as 35 kDa when analyzed by SDS-PAGE and western blotting. The rhLF-N was further confirmed by immunodiffusion using the anti-hLF polyclonal antibody. The expression profile analysis by qRT-PCR showed that the relative mRNA expression of rhLF-N was maximal after 2-3 days of methanol induction and reduced gradually at 4 days. The purified rhLF-N showed broad antibacterial activities against the pathogens such as Staphylococcus aureus, E. coli, Pseudomonas aeruginosa, Burkholderia cepacia, and Salmonella typhimurium. However, rhLF-N showed relatively lower activity when compared to peptides derived from LF. In spite of this weak activity, the rhLF-N expressed in P. pastoris might be more advantageous for the industrial application, because rhLF-N is secreted into the culture medium and the production can also be increased by optimization of culture conditions.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2000.11a
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• pp.182-195
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• 2000

Indigenous bacteria from poplar roots (Populus mnadensis var. eugenei, 'Imperial Carolina') and Southern Californian shrub rhizospheres as well as two tree-colonizing Rhizobium strains (ATCC 10320 and 35645) were genetically engineered to express constitutively and stably toluene o-monooxygenase (TOM) from Burkholderia cepacia G4 by integrating the torn locus into the chromosome. The poplar and Rhizobium recombinants degraded trichloroethylene (TCE) at 0.8-2.1 nmol/min.mg protein (initial TCE concentration, 10u M) and competitive against the unengineered hosts in wheat and barley rhizospheres for one month (colonization at 1-23 $\times$ 10$^{5}$ CFU/cm root). In addition, six of these recombinants colonized poplar roots stably and competitively with populations as high as 79 $\pm$ 12% of all rhizosphere bacteria after 28 days (0.2-31 $\times$ 10$^{5}$ CFU/cm root). Furthermore, five of the most-competitive poplar recombinants (e.g., Pb3-1 and Pb5-1 which were identified as Pseudomonas PsK) retained the ability to express TOM for 29 days as 100 $\pm$ 0% of the recombinants detected in the poplar rhizosphere had constitutive expression of TOM.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.370-373
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• 2000

In this work, an extractive membrane bioreactor containing coulture broth of Burkholderia cepacia G4 PR1 constitutively expressing the TCE-degrading enzyme, tolune-ortho-monooxygenase(TOM), was used for the degradation of TCE. The membrane bioreactor operates by seperating the TCE-containing waste gas from the aerated biomedium, by which the air-stripping of TCE without degradation was overcome that could occur in conventional aerobic biological treatments of TCE-contaminated waste gases. This was achieved by a silicone rubber membrane which was coiled around a perspex draft tube. TCE from the gas phase diffuses across the silicone rubber membrane into microbial culture broth that was continuously fed from a separate aerobic CSTR. Therefore, TCE degradation occured without the TCE being directly exposed to the aerating gas stream. Of the TCE supplied to the membrane bioreactor, 72.6% was biodegraded during the operation of this system. To construct a mathematical model for this system, parameters describing microbial growth kinetics on TCE were determined using a CSTR bioreactor. Else parameters used for numerical simulation were determined from either indepedent experiments or values reported in the literature. The model was compared with the experimental data, and there was a good agreement between the predicted and the measured TCE concentrations in the system. To achieve a higher treatment efficiency, various operating conditions were simulated as well.

• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.258-263
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• 2000

2-Hydroxy-6-oxo-6phenylhexa-2,4-dienoate (HOPDA) hydrolase catalyzes the hydrolytic cleavage of HOPDA to bemzpate and 2-hydroxypenta-2, 4-dienoate (HPD) during microbial catabolism of biphenyl and polychlorinated biphenyls. A HOPDA hydrolase gene (pcbD) was isolated from the genomic library of Pseudomonas sp. P20 and designated as pCNUO1201; a 7.5-kb XbaI DNA fragment from Pseudomonas sp. P20 was inserted into the pBluescript SK(+) XbaI site. E. coli HB101 harboring pCNU1201 exhibited HOPDA hydrolase activity. The open reading frame (ORF) corresponding to the pcbD gene consisted of 855 base pairs with an ATG initiation codon and a TGA termination codon. The ORF was preceded by a rebosome-binding sequence of 5'-TGGAGC-3' and its G+C content was 55 mol%. The pcbD gene of Pseudomonas sp. P20 was located immedeately downstream of the pcbC gene encoding 2,3- dihydroxybiphenyl 1,2-dioxygenase, and approximately 4-kb upstream of the pcbE gene encoding HPD hydratase. The pcbK gene was able to encode a polypeptide with a molecular weight of 31,732 containing 284 amino acid residues. The deduced amino acid sequence of the HOPDA hydrolase of Pseudomonas sp. P20 exhibited high identity (62%) with those of the HOPDA hydrolases of P. putida KF715, P. pseudoalcaligenes KF707, and Burkholderia cepacia LB400, and also significant homology with those of other hydrolytic enzymes including esterase, transferase, and peptidase.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.22 no.4
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• pp.420-429
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• 2000

Oral & maxillofacial infections are most commonly odontogenic in origin. Although such infections are usually self-limiting, they may occasionally spread deeply into fascial spaces or planes far from the initial site of involvement. If early diagnosis and appropriate therapy is delayed, complications such as mediastinal extension, retropharyngeal spread and airway obstruction could happen to the patients. For the study of the microbiology, we have retrospectively analysed the oral & maxillofacial infected patients in the Dept. of Oral & Maxillofacial Surgery. In-Ha University Hospital from 1997 September to 2000 April. The results were as follows 1. The male patients were more common than female, with male 61.9% and female 38.1%. 2. Dental originated infections were most common cause with the incidence of 62%. 3. Most common fascial space involved was buccal space 42cases(37.2%) followed by submandibular space 13cases(11.5%), infraorbital space 13cases(11.5%), masseteric space 11cases(9.7%), periapical abscess 11cases(9.7%). 4. The causative organisms isolated from the pus culture were Gram Positive Bacterial species, which were 46cases(31.9%) of Streptococcus viridans, 16cases(8.6%) of ${\alpha}$ and ${\beta}-hemolytic$ streptococcus, 4cases(3.1%) of Strep.-group D non enterococci, 7cases(5.1%) of Staphylococcus Coa. neg., 5cases(3.9%) of Staphylococcus aureus, 3cases(2.3%) of Enterococcus faecalis, 1case(0.8%) of Bacillus species, 1case(0.8%) of Peptostreptococcus, 1case(0.8%) of Clostridium and Gram negative bacterial species, which were 4cases(3.1%) of Acinetobacter baumannii, 2cases(1.6%) of Pseudomonas aeruginosa, 2cases(1.6%) of Burkholderia cepacia, 1case(0.8%) of Neisseria species, 1case(0.8%) of Klebsiella pneumoniae, 1case(0.8%) of Klebsiella oxytoca, 1case(0.8%) of Escherichia coli. 5. In drug sensitivity test, high resistant tendency was found in Penicillin system(Penicillin G 83.3%, Ampicillin 60%) and Aminoglycosides (Gentamycin 50%, Tobramycin 45.5%), but tertiary Cephalosporin system(Cefoperazone 9.1%, Ceftazidime 18.2%), and glycopeptides system (Teicoplanin 0%, Vancomycin 0%) showed lower resistancy.