Being autotrophic, algae occupy a trategic place in the biosphere. They produce oxygen both directly and indirectly through the chloroplasts of all green plants. The chloroplasts are believed to have originated from archaic prokaryotic algae through endosymbiosis with primitive eukaryotic cells. Phytoplankton and other algae regulate the global environment not only by releasing oxygen but also by fixing carbon dioxide. They affect water quality, help in the treatment of sewage, and produce biomass. They can be used to produce hydrogen which is a clean fuel, and biodiesel, and fix $N_2$ for use as a biofertilizer. Some other services of algae to the environment include restoration of metal damaged ecosystems, reducing the atmospheric $CO_2$ load and citigating global warming, reclamation of saline-alkaline unfertile lands, and production of dimethyl sulphide (DMS) and oxides of nitrogen (NOx) involved in the regulation of UV radiation. ozone concentration, and global warming. Algae can be valuable in understanding and resolving certain environmental issues.

Environmental stres is known to induce heat shock proteins (HSPs) in eukaryotic cells. However, the induction of HSPs in host cells by microbial infection has not yet been well explained. Staphylococcus aureus (S. aureus) is one of the major pathogens in the pathogenesis of endovascular diseases such as infective endocarditis. In this study, the synthesis of stress-inducible 70 kDa HSP was investigated in the endothelial cells (ECs) after 3 h to 20 h of incubation with S. aureus. The dffect of S. aureus infection on the expression of HSP70 in cultured ECs was analyzed using laser scanning confocal microscopy (LSCM). The increase of HSP70 expression in ECs infected by S. aureus ($10^4{\;}cfu/ml$) for 20 h was 1.1-fold higher than that in heat shock treated ECs and 2.2-fold higher than that in untreated cells. Heat shock is known to induce intranucleus HSP70 expression in mammalian cells, whereas the S. aureus infection induced perinuclear expression in ECs as observed by LSCM. Consequently, the expression of HSP70 in ECs plays an important role in the pathogenesis of endovascular infection.

This study was conducted to compare the cellular fatty acid composition and genotypic analysis of Bifidobacterium longum MK-G7 originated from Koreans with other commercial type strains of bifidobacteria. The cellular fatty acid of Bif. longum MK-G7 was shown to be composed of $C_{160FAME},C_{181\;c18DMA},C_{18.1\;CIS9\; FAME},C_{14.0FAME},C_{19\;0cye9,10 DMA},Feature7(C_{17.2 FAME), and Feature 10(C_{181\; Cll/t9/t6 FAME}$. Bif. longum MK-G7 showed 99.9% homology and the highest relatedness with Bif. longum ATCC 15707 type strain. Both Bif. longum MK-G7 and Bif. longum ATCC 15707 showed 153 bp products on RAPD (randomly amplified polymorphic DNA) analysis, however, they showed quite different band patterns on PFGE (pulsed-field gel electrophoresis) analysis. Consequently, our present study showed that Bif. longum MK-G7 was different from any commercial type strains of Bif. longum tested.

This study was conducted to compare probiotic activities and physiological functions of Bifidobacterium longum Mk-G7 with weveral commercial and type strains of bifidobacteria. bif. longum MK-G7 showed the highest acid tolerance against HCl and acetic acid, whereas bif. infantis Y-1 showed the lowest acid tolerance and more than 4 log cycles of viable cell count decreased due to acid injuty. Viable cell counts of bifidobacteria strains decreased more than 1.5 log cycles owing to oxygen toxicity, with the exception of Bif. longum MK-G7, Bif. infantis Y-2, Bif. longum Y-3, Bif. longum Y-6, and Bif. longum RD-13 showed the highest bile tolerance, whereas Bif. longum MK-G7 showed a medium level of bile tolerance. Only Bif. longum MK-G7 howed much higher antibiotic resistance against both tetracycline and penicillin-G in the MIC(minimum inhibitory concentration) level of 24.8 mg/I and 0.52mg/I, respectively. Bif longum Y-6, and Bif. bifidum ATCC 29539 showed more than 80% of anti-mutagenicity against NQO(4-nitroquinolinel-oxide). Since the production of cytokines such as $TNF(tumor necrosis factor)-{\alpha}$ and IL (interleukin)-6, and NO(nitric oxide) in the macrophage cell line Raw 264.7 cells increased as Bif. longum MK-G7 cell concentration increased, ti was suggested that Bif. longum MK-G7 is able to enhance immunopotentiating activity in vitro. When freeze-dred Bif. longum MK-G7 was administered to mice at the dose of 1,2,4, and 6 g/kg of body weight, all of the mice survived in all feeding groups, proving the GRAS(generally recognized as safe) status of Bif. longum MK-G7. When fermented milk containing Bif. longum MK-G7 was administered to human volunteers, viable cell count of total bifidobacteria and anaerobes in the feces increased up to 0.5 log cycles more than before the administration. In particular, Bif. logum MK-G7 ingibited the growth of Bacteroides at the level of 1.0-1.5 log cycles.

Aflatoxin B1 is known as the most potent mycotoxin produced by several fungi. It has been demonstrated to be not only carcinogenic but teratogenic and mutagenic as well in humans. To prevent or inactivate aflatoxins, several chemical of physical methods were tested for ammoniation, using insecticides as an wxample, but they were unsuitable for food products. On the contrary, biological control by antagonistic microorgani는 is and ideal method. In order to control aflatoxin B1 biologically, the antagonists #07, #63, #75, #74, and #61 were separated from various samples by using the antagonistic activity test. Among them, culture filtrate part A (non heat-treated) of #63 and #74 on aflatoxin B1 produced by Aspergillus fkavus were shown to be 95% and 75%, respectively. Based on the morphological characteristics, #63 was deduced as an Azospirillum sp.

Plasmid pLEX3 isolated from the recombinant cosmid library of Zoogloea ramigera 115 was found to be responsible for the restoration of the rugose colony phenotype. To confirm the essential region responsible for the complementation, subclones were constructed from plasmid pLEX3 and transformed into mutant strain Z. ramigera 115SLR. The recombinant plasmids pLEX10 and pLEX11 were shown to complement the slime-forming property of Z. ramigera 115SLR. In a compositional analysis of the exopolysaccharides from Z. ramigera 115, Z. ramigera 115SLR, and Z. ramigera 115SLR harboring plasmid pLEX11, the exopolysaccharides showed a similar composition with glucose, galactose, and side chain groups. The complete nucleotide sequence of the 3.25kb genocim DNA insert in plasmid pLEX11 was determined and its analysis identified two open reading frames which could encode two proteins. The gene products derived form the two open reading frames were confirmed by and in vivo transcription using a T7-RNA polymerase. The ORF1 produced a 30 kDa protein, whereas the ORF2 was found responsible for the complementation of the morphological mutation and produced a 14 kDa protein. An in vivo gene expression of plasmid pTEX10 showed another open reading frame encoding a 50 kDa protein. The gene products form ORF1 and ORF2 are regarded as novel proteins which do not show any homology with other proteins.

The filamentous soil bacterium Streptomyces coelicolor is known to produce four distinct antibiotics. The simultaneous global regulation for the biosynthesis of those four antibiotics was previously confirmed by absA and absB mutations that blocked all four antibiotics' biosynthesis without influencing their morphological differentiation. To study the complex regulatory cascade that controls the secondary metabolism in Streptomyces, a new abs-like mutation was characterized. namely absR, which is slightly leaky on a complete R2YE medium, yet tight on a minimal medium. A genetic analysis of the absR locus indicated that it is located at 10 o'clock on the genetic map, near the site of absA. A cloned copy of the absA gene that encoded bacterial two-component regulatory kinases did not restore antibiotic biosyntheis to the absR mutant. Accordingly, it is proposed that absR is another abs-type mutation which is less tight than the previously identified absA or absB mutations income medium conditions, and can be used to characterize another global regulatory gene for secondary metabolete formation in S. coelicolor.

Three polyene macrolide antibiotics including two new compounds were isolated from the culture mycelia of a Streptomyces species. The structures of these metabollites were determined as elizabethin, a previously reported 28-membered macrolide and two analohs, using combined spectroscopic methods. These compounds exhibited antifungal activity and cytotoxicity against a juman leukemia cell.

MAPI (microbial alkaline protease inhibitor) was isolated from cultrue broth of Streptomyces chromofuscus SMF28. The Ki values of MAPI for the representative serine proteases such as chymotrypsin and proteinase K were 0.28 and $0.63{\;}\mu\textrm{M}$, respectively, and for the cysteine proteases cathepsin B and papain were 0.66 and $0.28{\;}\mu\textrm{M}$, respectively. These data indicate that MAPI is not a potent selective inhibitor of serine or cysteine proteases. Progress curves for the inhibition of three proteases by MAPI exhibithe characteristic patterns; MAPI exhibited slow-binding inhibition of cathepsin B. It was rapidly associated with chymotrypsin before the addition of substrate and then reactivation of MAPI-inhibited enzyme was investigated in the presence of substrate. On the other hand, MAPI-proteinase K interaction was typical for those classical inhibitors. When MAPI was incubated with trypsin, there was an extensive reduction in the ingibitory activities of MAPI corresponding to 66.5% inactivation of MAPI, indicating that trypsin-like protease may play a role in the decrease of the inhibitory activity during cultivation.

An actinomycetes strain, T-2, which produces transglutaminase (EC 2.3.2.13), was isolated from soil and identified as belonging to the Actinomadura sp., based on taxonomc studies. The conditions for the transglutaminase production and its enzymatic properties were investigated. The optimum components for the transglutaminase production were 2% glucose, 1% polypeptone and soytone, and 0.1% MnCl2. The optimum pH and temperature of the enzyme reaction were pH 8.0 and $45^{\circ}C$, respectively. The enzyme was stable within the pH range of 5.0-9.0 and $30^{\circ}C-45^{\circ}C$. The novel enzyme required no calcium ions for its activity. This enzyme polymerized various proteins such as casien, soy protein, hemoglobin, egg white, gelatin, and soybean milk.

Bacillus cereus BS229 was identified as a bacteriocin producer with a bactericidal activity against Bacillus thuringiensis subsp. Thomsoni BR-40. Bacillus cereus BS229 and cerein BS229, named tentatively as the bacteriocin produced by Bacillus cereus BS229, showed a narrow spectrum of actibity against Gram-positive and Gram-negative bacteria, along with yeast and molds. Production of cerein BS229 in a 5-1 fermenter followed typical kinetics of primary metabolite synthesis. The antibacterial activity of cerein BS229 on sensitive indicator cells disappeared completely by ${\alpha}-chmotrypsin$ or proteinase K, which indicates its proteinaceous nature. Cerein BS229 seemed to be very stable throughout the pH range of 2.0 of 9.0 and it was relatively heat labile, despite the fact that bacteriocin activity was still detected after being boied for 30min. Cerein BS229 actibity has been changed with some of the organic solvents such as toluene, ethanol, and chloroform. Direct detection of cerein BS229 actibity on SDS-PAGE suggested that it had an apparent molecular mass of about 8.2 kDa.

The freshwater green alga Chlorococcum sp. grew on NH_4^{+},{\;}NO_3^{-}$, urea, yeast extract, and peptone as the nitrogen source showing similar pattens of growth and secondary carotenoid (SC) production. However, the most suitable nitrogen source for the induction fo SC was urea. The dffects of nutrient levels (urea, phosphate, sulfate, ferrous iron, and salt) on growth and SC production were stydied by varying the concentration of each nutrient in batch cultures. High biomass production was achieved in cultures containing 20-28 mM urea, 4.8-10 mM phosphate, 1.6 mM sulfate, 70 mM phosphate, 1.6 mM sulfate, 170 mM NACl, and $50{\;}\mu\textrm{M}$ iron. The optimum concentrations of nutrients for biomass and for the SC accumulation in biomass were evaluated and the two media for achieving high biomass production and SC production were thus developed. The extent to which each parameter to stimulate the formation of SC in the alga were varied and the potentially improned SC prodution by manipulating the nutrient levels in the modified media were descussed.

A bacterial strain producing extracellular chitosanase was isolated from crab shells and identified as a member of the genus Aureobacterium The production of chitosanase was proportionally related to the microbial growth, induced by the presence of chitosan, and repressed by glucose at 0.5% (w/v) concentration or higher. The optimal culture conditions for the production of chitosase were 3$0^{\circ}C$ and pH 7.0. Among the nitrogen sources tested, incubation with 0.25% (w/v) concentrations of tryptone and casitione showed the best production of chitosanase. The chitosanase of Aureobacterium sp. YL produced chitobiose as a major product and glucosamine, chitotriose, chitotetraose, and chitopentaose as minor products from chitosan.

Abstract White spot disease (WSD), resulting in more than 90% mortality of aquacultured penaeid shrimp, has been reported off the southern and western coasts of Korea since 1993. The pafuogen of WSD has been identified as being a virion wifu an envelope around a central nucleocapsid, and with an average size of 167 nm in diameter and 375 nm in length. In the present study, an in situ hybridization technique was developed as a rapid. sensitive, and specific diagnostic assay for the WSD viros infection in shrimp. Furthermore. the pathological changes ofWSD, in shrimp experimentally infected with WSD viroses. were investigated. Using a biotinylated 643 bp probe obtained from a peR using primers specific to the rod-shaped virus of Penaeus japonicus (RV-PJ), positive signals were detected in both naturally and experimentally infected shrimps. The in situ hybridization revealed positive reactions in the nuclei of the stromal matrix cells in the lymphoid organ, epithelia of the gills, foregut. epidermis, and hematopoietic cells of the interstitial tissues, suggesting the presence of WSD virus. Tills result indicates that the in situ hybridization method can be useful for a rapid and sensitive detection of WSD viruses in shrimp.shrimp.

Cephalosporin-C deacetylase (CAH) catalyzes the deacetylation of cephalosporin derivatives. A novel gene encoding the CAH from Bacillus sp. KCCM10143 was cloned and sepuenced. The uncleotide sequence contained an open reading frame encoding a polypeptide consisting of 217 amino acids and a molecular weight of 24 kDa which was in good agreement with the value obtained by sodium dodecylsulfate-polyacrylamide gel electrophoresis. An expression plasmid was constructed by inserting the CAH gene into the region of the pTrc99A expression vector. An active from of the CAH protein was expressed in the soluble fraction obtained after cell disruption. in fermentation using a 5-1 jar fementer, the transformant E. coli JM109 (pDST654) produced 4.12 U of CAH per ml of culture during 16 h of incubation.

Fresh pine needles were collected and extracted with 95% methanol and the extract was concentrated to determine its antimicrobial activity. The methanol extract had a considerable inhibitory effect on the tested bacteria, such as Esherichia coli 0157;H7, Listeria monocytogenes, Salmonella typhimurium, and Staphylococcus aureus. The methanol extract of pine needles was further fractionated to chloroform, ethylacetate, butanal, and water fractions. Among these four fractions, the butanol and water fractions, which showed a relatively strong inhibitory effect on all of the tested bacteria, were purified and the minimum ingibitory concentration (MIC) was determined for each microorganism. The MIC raged between 25 mg/ml and 45 mg/ml depending on the microorganism. the purified active fractions were applied to sterilized milk as a model food system to define the antimicrobial effectiveness and it was found that the antimicrobial activities in the water fractions were stronger than those in the butanol fractions.

As part of a continuing investigation to identify free radical scavengers from the fruit bodies of basidiomycetes, we isolated two p-terphenyl compounds, designated as PAl and PA2l, from cethanolic extract of the fruit body of Pasillus panuoides. The methanolic extract was processed by ethyl acetate extraction and silica gel column chromatography to yield two active fraction. PAl was obtained from one of the fractions through Sephadex LH-20 and silica gel column chromatographies and reverse-phase HPLC. The other fraction was purified by Sephadex LH-20 and reverse-phase column chromatographies to produce PA2. The compounds PA1 and PA2 were identified as leucomentin-4 and leucomentin-2, respectively, on the basis of various spectroscopic analyses. These compounds exhibited strong inhibitory activities against lipid peroxidation in rat liver microsomes with IC_{50}$ values of 0.10 and $0.06{\;}\mu\textrm{g}ml$, respectively.

An exoinulinase (${\beta}-D-fructofuranosidase$) gene was cloned by chromosome walking along the upstream region of the endoinulinase gene of Pseudomonas mucidolens isolated from soil. the exoinulinase gene consisted of an ORF of 0,506 bp encoding a polypeptide of 501 amino acids with a deduced molecular weight of 55,000. The exoinulinase produced by the recombinant Escherichia coli $DH5{\alpha}$ strain was also purified to homogeneity as determined by SDS-PAGE and a zymogram. The molecular weight of the purified exoinulinase according to both SDS-PAGE and gel filtration matched the deduced molecular weight of the protein described above, thereby indicating that the native form of the exoinulinase was a monomer. The purified enzyme hydrolyzed activity value of 2.0. Furthermore, no inulo-oligomers were liberated from the inulin substrate in the enzymatic reaction mixtures incubated for 90 min at $55^{\circ}C$. Taken together, these results indicate that the purified ${\beta}-D-fructofuranosidase$ was an exoinulinase. The pH and temperature optima of the exoinulinase were pH 6.0 and $55^{\circ}C$, respectively. the enzymehad no apparent requirement for a cofactor, and its activity was completely inactivated by $Ag^{+},{\;}Hg^{2+},{\;}and{\;}Zn^{2+}$. Kinetic experiments gave $K_m,{\;}V_{max},{\;}and{\;}K_{cat}$ values for inulin of 11.5 mM, 18 nM/s, and $72{\;}s^{-1}$, respectively. the exoinulinase was fairly stable in broad pH conditions (pH 5-9), and at pH 6.0 it showed a residual activity of about 70% after 4 h incubation at $55^{\circ}C$.

An ${\alpha}-glucosidase$ gene (aglA) from thermophilic Bacillus sp. DG0303 was cloned, sequenced, and expressed in Escherichia coli. The aglA was localized to the 2.1-kb PvuI-XmnI region within the 5.9-kb DNA insert of the gybrid plasmid pAG1. The gene consisted of an open reading frame of 1,686 bp with an unusual GTG initiation codon and TGA termination codon. The amino acid sequence deduced from the nucleotide sequence predicted a protein of 562 amino acid residues with a M, of 66,551 dalton. A comparative amino acid sequence analysis revealed that DG0303 ${\alpha}-glucosidase$ is related to bacillary oligo-1, 6-glucosidases. The Bacillus sp. DG0303 ${\alpha}-glucosidase$ showed a high sequence identity (36-59%) to the B. flavocaldarius, B. cereus, and B. thermoglucosidasius oligo-1, 6-glucosidases. The number of prolines in theses four ${\alpha}-glucosidases. was observed to increase with increasing thermostability of these enzymes. The cloned ${\alpha}-glucosidase was purified from E. coli $DH5{\alpha}$ bearing pAG1 and characterized. The recombinant enzyme was identical with the native enzyme in its optimum pH and in its molecular mass, estimated by sodium dodecy1 sulfate-polyacrylamide gel electrophoresis. The temperature optimum of the cloned ${\alpha}-glucosidase$ was lower than that of the native enzyme.

Chitin synthases are identified as key enzymes of chitin biosynthesis in most of the fungi. Among them, chitin synthase II has been reported to be and essential enzyme in chitin biosynthesis, and exists as a membrane-bound form. To search and screen new antifungal agents from natural resources to inhibit chitin synthase II, the assay conditions were established using the enzyme isolated from Saccharomyces cerevisiae ECY38-38A(pAS6) that overproduces only chitin synthase II. This enzyme was activated only by partial proteolysis with trypsin. Its actibity reached the maximum at $80{\;}\mu\textrm{g}/ml$ of trypsin and was strongly stimulated by 2.0 mM $Co^{2+}$, 1.0 nM UDP-[$^{14}C$]-GicNAc, and 32 mM free-GlcNAc. Under these assay conditions, the highest chitin synthase II activity was observed by incubation at $30^{\circ}C$ for 90 min. However, and extremely narrow range of organic solvents up to as much as 25% of DMSO and 25% of MeOH was useful for determining optimal assay conditions. After a search or potent inhibitors of chitin synthase II from natural resources, prodigiosin was isolated from Serratia marcescens and purified by solvent extration and silica gel column chromatographies. The structure of prodigiosin was determined by UV, IR, Mass spectral, and NMR spectral analyses. Its molecular weight and formula were found to be 323 and $C_{20}H_{25}N_{3}O$, respectively. Prodigiosin ingibited chitin synthase II by 50% at the concentration of $115{\;}\mu\textrm{g}/ml$.

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.

A new GFP-based T-vector for cloning of PCR products was developed by using a green fluorescent protein (GFP) as a mafker. In order to facilitate the DNA inserts, multiple restriction sites, SP6 and T7 RNA polymerase promoter sites, were introduced close to the PCR DNA insertion site of a pCRGv vector. The XcmI-digested pHNT plasmid can be used to clone a 3' A-overhanged PCR DNA amplified by Taq DNA polymerase. A potential method of easing some difficulties from its use along with its cost savings proveded by this vector are likely to lead to the replacement of other T-vectors for PCR DNA cloning.

Abstract Two glucans (PONGa and PONGb) differing in their anomeric and glycosidic linkage structures were isolated from the water-insoluble materials (PON) of Pleurotus ostreatus basidiocarps, which activated the complement system and were almost soley composed of D-glucose. The isolatIon was achieved by repeated precipitations with ethanol and adsorption on concanavalin A (Con A) of paN suspension in thymol/NaCL Based on methylation analysis. IR, GLC-MS, $^1H,{\;}and{\;}^{13}C-NMR$ spectroscopies, PONGa was found to be a branched a-glucan composed of ${\alpha}-linked$ D-glucopyranose residues and ${\alpha}-linked$ units with 6-branching points, whereas PONGb was a linear ${\beta}-1,3-glucan$ composed mainly of ${\beta}-1,3-linked$ D-glucopyranose residues. The PONGb particles reacted more potently than the PONGa particles as C3 activator in alternative complement hemolysis and crossed-immunoelectrophoresis using anti-human C3, thereby suggesting that the complement activating components of PON were ${\beta}-(13)-glucans rather$ than ${\alpha}-glucan$ components.onents.

Temperate bacteriophage P2 has a nonessential gene called old(overcoming lysoginization defection). In the presence of old, the growth of the host (Escherichia coli) with recBC- genotype is ingibited, and another bacteriophage, lambda, cannot superinfect. The Old protein has been shown to possess an exonuclease actibity. Three mutant P2s(old 1, old 17, old 49) which did gene was coned into expression vectors to produce hexahistidine-tagged proteins. The proteins were affinity-purified and shown to lose its exonuclease activity on both double-stranded and single-stranded DNA substrates. Thus, it was concluded that the lambda exclusion was related to Old's exonuclease activity.

The inulin fructotransferse (depolymerizing) (IFTase, EC 2.4.1.93) gene of Arthrobacter sp. A-6 was cloned and expressed in Escherichia coli and Bacillus subtilis. The IFTase gene consisted of an ORF of 1.311 nucleotides encoding a polypeptide of 436 amino acids containing a signal peptide of 31 amino acids in the N-terminus. The molecular mass of the IFTase based on the nucleotide sequence was calculated to be 46.116 Da. The recombinant E. coli $DH5{\alpha}$ cells expressing the Arthrobacter sp. A-6 IFTase gene produced most of the IFTase intracelularly. In contrast, the recombinant B. subtilis DB 104 carrying the IFTas gene on a B. subtilis-E. Coli expression vector secreted the IFTase into the culture fluid efficiently.