• Korean Journal of Microbiology
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• v.37 no.2
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• pp.158-163
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• 2001

The effect of phosphate on the production of avermectin B1a was studied. Response surface methodology was applied to optimize the concentration of organic nitrogen sources. The portion of B1b in total avermectins was decreased from 5.8% to 3.0% by the addition of 1.5 g/ι inorganic phosphate to the production medium. Among organic nitrogen sources, soybean meal was the most effective on avermectin biosynthesis. Results showed that B1a productivity was increased by 44.8% in a laboratory scale fermenter cultivation of Streptomyces avermitilis YA99-40 through fed-batch process. A maximal B1a productivity was obtained by repeated 30 and 20 g/ι of glucose feeding at 136 and 206 hour, respectively. The B1a productivity was increased by 86.3% and the proportion of B1a in the total avermectins was improved from 38% to 45% with respect to the control process. These results would be very useful for enhancing productivity of B1a in an up-scaled processes.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.152-156
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• 2005

In this study, we tried to optimize the composition of medium and culture conditions for the total avermectin and avermectin B1 production from S. avermitilis, which is a natural producer of avermectin family. Among various carbon and organic nitrogen sources tested, fructose and malt extract were most effective on avermectin production. Next addition of polyethylene glycol and $K_{2}HPO_{4}$ in medium significantly improved the intracellular contents of avermectin. Thus the optimized medium composition was 50 g/L fructose, 30 g/L malt extract, 5 g/L casamino acid, 2.5 g/L PEG 3,350, and 1 g/L $K_{2}HPO_{4}$, which increased the avermection production from10 to 478 mg/L. The contents of avermectin B1 complex was about 50% of the total amount of avermectin.

• Applied Biological Chemistry
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• v.35 no.2
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• pp.115-121
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• 1992

Avermectin B1(abamectin) is an very effective acaricide/insecticide. Because it is applied at low rates, the resultant residue level would be quite low and this requires highly sensitive analytical method. In this study, three analytical methods for avermectin $B1_a$, were compared in view of detectability and sensitivity. The first analytical method was an HPLC method employing the fluorescence detection of avermectin. The second analytical technique to quantitate avermectin $B1_a$, and its photodegradative delta 8,9-isomer employed trifluoroacetic anhydride and 1-methylimidazole in DMF. The new method was the modification of trifluorescence method. The average recoveries of avermectin $B1_a$ for the concentration range from 1 and 10 ng/g in whole apple fruit by fluorescence method were 90.3% and 88.2% respectively. In trifluorescence method, the recoveries of the avermectin $B1_a$ and delta 8,9-isomer were 100.7% and 94.7% in concentration from 5 ng/g and 25 ng/g. The average recoveries of 5 ng and 25 ng/g in the newly modified method were 95.0, 99.0. 96.0, 92.8% in whole apple and soil respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.3 no.4
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• pp.337-341
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• 1990

An investigation was conducted to determine whether the rumen modifiers lasalocid and avoparcin, when included in molasses/urea based supplements, enhanced liveweight performance, in early weaned calves. As part of the study the broad-spectrum parasiticle Avermectin B1 was given to the calves to assess any undesirable side effects on animals of less than four months of age. There were no significant (p>0.05) liveweight responses to supplementation when the rumen modifiers lasalocid and avoparcin were included in supplement rations. Lasalocid reduced supplement intake, however, it had no adverse effect on liveweight gain. Avoparcin substantially improved growth when cottonseed meal was included in the ration. Weaners treated with Avermectin B1 tended to show a greater liveweight gain than untreated weaners during the experiment (p<0.10) and no adverse side effects were noted.

• Journal of Microbiology
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• v.43 no.3
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• pp.277-284
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• 2005

The avermectins are composed of eight compounds, which exhibit structural differences at three positions. A family of four closely-related major components, A1a, A2a, B1a and B2a, has been identified. Of these components, B1a exhibits the most potent antihelminthic activity. The coexistence of the '1' components and '2' components has been accounted for by the defective dehydratase of aveAI module 2, which appears to be responsible for C22-23 dehydration. Therefore, we have attempted to replace the dehydratase of aveAI module 2 with the functional dehydratase from the erythromycin eryAII module 4, via homologous recombination. Erythromycin polyketide synthetase should contain the sole dehydratase domain, thus generating a saturated chain at the C6-7 of erythromycin. We constructed replacement plasmids with PCR products, by using primers which had been derived from the sequences of avermectin aveAI and the erythromycin eryAII biosynthetic gene cluster. If the original dehydratase of Streptomyces avermitilis were exchanged with the corresponding erythromycin gene located on the replacement plasmid, it would be expected to result in the formation of precursors which contain alkene at C22-23, formed by the dehydratase of erythromycin module 4, and further processed by avermectin polyketide synthase. Consequently, the resulting recombinant strain JW3105, which harbors the dehydratase gene derived from erythromycin, was shown to produce only C22,23-unsaturated avermectin compounds. Our research indicates that the desired compound may be produced via polyketide gene replacement.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.101-109
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• 2004

In order to enhance the productivity of AVM $B_{la}$ produced by Streptomyces avermitilis as a secondary metabolite, we established a basic protocol necessary for protoplast fusion with high-producing strains as a fusion partner, and then obtained various kinds offusants by adopting a massive strain-development procedure (a miniaturized strain screening system). An alternative fusion method using UV and/or NTG mutation of protoplasts was developed to screen genetic recombinants without specific selectable markers. In this method, the mutants obtained by protoplast fusion after UV and/or NTG treatment (95% death rate) of the respective fusion partner (protoplasts of the respective mutants resistant against L-isoleucine antimetabolites such as O-methylthreonine and/or azaleucine) were regarded as DNA-recombined protoplast fusants. Notably it was demonstrated that most of the protoplast recombinants obtained by the UV mutation method were able to biosynthesize higher amount of AVM $B_{la}$ , reaching almost three times higher level (almost equal to the industrial productivity), compared to the average AVM Bla amount of the parallel mother strains.

• KSBB Journal
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• v.20 no.5 s.94
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• pp.376-382
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• 2005

Avermectin (AVM) $B_{1a}$ produced by Streptomyces avermitilis via polyketide pathway is a secondary metabolite with powerful anthelmintic and insecticidal activities, thus being used as an efficient agent in the field of agriculture and animal health. It has been reported that a precursor for AVM $B_{1a}$ biosynthesis was isoleucine and the biosynthetic pathway of AVM $B_{1a}$ was closely similar to that of fatty acid. Based on understanding of the biosynthetic pathway of AVM $B_{1a}$, we intended to screen various mutants resistant against O-methyl threonine (OMT), an isoleucine-anti metabolite, and/or mutants resistant against p-fluoro phenoxy acetic acid (pFAC), an inhibitor of fatty acid biosynthesis. It was inferred that these mutants could produce AVM $B_{1a}$ more efficiently, due to the acquired capability of not only overproducing isoleucine intracellularly but also channelling metabolized carbon-sources into the polyketide pathway, thus leading to enhanced biosynthesis of AVM $B_{1a}$. The resulting mutant (PFA-1 strain) resistant against 100 ppm of pFAC was able to produce approximately 42 fold higher amount of AVM $B_{1a}$ compared to the parallel mother strain (4,200 vs. 100 units/l). In addition, through the process of continuous strain improvement program carried out by gradually increasing the OMT concentration, it was possible to obtain a more attractive mutant with greater AVM $B_{1a}$ production capacity (9,000 units/l). Notable was that significantly higher producer (12,000 units/l) could be selected through further screening of the resistant mutants, this time, to even higher concentration of PFAC. Meanwhile, through the analysis of AVM Bla production histograms (i.e., number of strains according to their AVM $B_{1a}$ biosynthetic ability) for the earlier strains in comparison with the high producers having the characteristics of resistance to OMT and pFAC, it was found that production stability of the high-yielding producers were remarkably improved, as demonstrated by the fact that larger proportion of the mutated strains had greater capability of AVM $B_{1a}$ biosynthesis ($71\%$ in the range between 5,000 and 7,000 units/L; $47\%$ in the range between 6,000 and 7,000 units/l). Based on these consequences, it was concluded that the rational screening strategy based on the understanding of the biosynthetic pathway of AVM $B_{1a}$ was very effective in obtaining high-yielding mutants with the features of enhanced production stability.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.135-135
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• 2005

• Proceedings of the Zoological Society Korea Conference
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• 2000.10a
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• pp.287.1-287
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• 2000

No Abstract, See Full Text

• Journal of Microbiology and Biotechnology
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• v.16 no.11
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• pp.1690-1698
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• 2006

In order to investigate the effect of dissolved oxygen (DO) level on AVM $B_{1a}$ production by a high yielding mutant of Streptomyces avermitilis, five sets of bioreactor cultures were performed under variously controlled DO levels. Using an online computer control system, the agitation speed and aeration rate were automatically controlled in an adaptive manner, responding timely to the oxygen requirement of the producer microorganism. In the two cultures of DO limitation, the onset of AVM $B_{1a}$ biosynthesis was observed to casually coincide with the fermentation time when oxygen-limited conditions were overcome by the producing microorganism. In contrast, this phenomenon did not occur in the parallel fermentations with DO levels controlled at around 30% and 40% throughout the entire fermentation period, showing an almost growth-associated mode of AVM $B_{1a}$ production: AVM $B_{1a}$ biosynthesis under the environments of high DO levels started much earlier than the corresponding oxygen-limited cultures, leading to a significant enhancement of AVM $B_{1a}$ production during the exponential stage. Consequently, approximately 6-fold and 9-fold increases in the final AVM $B_{1a}$ production were obtained in 30% and 40% DO-controlled fermentations, respectively, especially when compared with the culture of severe DO limitation (the culture with 0% DO level during the exponential phase). The production yield ($Y_{p/x}$), volumetric production rate (Qp), and specific production rate (${\bar{q}}_p$) of the 40% DO-controlled culture were observed to be 14%, 15%, and 15% higher, respectively, than those of the parallel cultures that were performed under an excessive agitation speed (350 rpm) and aeration rate (1 vvm) to maintain sufficiently high DO levels throughout the entire fermentation period. These results suggest that high shear damage of the high-yielding strain due to an excessive agitation speed is the primary reason for the reduction of the AVM $B_{1a}$ biosynthetic capability of the producer. As for the cell growth, exponential growth patterns during the initial 3 days were observed in the fermentations of sufficient DO levels, whereas almost linear patterns of cell growth were observed in the other two cultures of DO limitation during the identical period, resulting in apparently lower amounts of DCW. These results led us to conclude that maintenance of optimum DO levels, but not too high to cause potential shear damage on the producer, was crucial not only for the cell growth, but also for the enhanced production of AVM $B_{1a}$ by the filamentous mycelial cells of Streptomyces avermitilis.