• Journal of Plant Biotechnology
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• 제5권2호
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• pp.121-129
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• 2003

Callus cultures of Azadirachta indica flower petals were established on MS medium supplemented with naphthalene acetic acid (1 mg/L), kinetin (0.5 mg/L) and sucrose $(3\%\;w/v)$. Cell cultures of Azadirachta indica were established and studied the growth and production kinetics. Half 85 medium supplemented with dicamba (2 mg/L), kinetin (1 mg/L) and sucrose $(3\%\;w/v)$ was found to be suitable for initiation and maintenance of cell cultures from the calli. MS medium supplemented with naphthalene acetic acid (NAA) (1 mg/L), kinetin (0.5 mg/L) and sucrose $(3\%\;w/v)$ was found to be suitable as production medium. Around $80\%\;(0.05\%\;w/v)$ of azadirachtin was found to be intracellular. The effect of various precursors, elicitors, permeabilizing agents and growth retardants in cell cultures was studied. The addition of precursors sodium acetate (10 mg/L), squalene (10 mg/L), isopentenyl pyrophosphate (1 mg/L) and geranyl pyrophosphate (1 mg/L) to the cell cultures on day 3 has shown significant increase in bioproduction of azadirachtin $(64.94{\pm}4.40\;mg/L,\;72.81{\pm}0.04\;mg/L,\;51.63{\pm}1.26\;mg/L\;and\;30.70{\pm}0.28\;mg/L\;respectively)$ over the control cultures $(4.70{\pm}0.27 mg/L)$. $5\%$ v/v cell extracts of Fusarium solani has shown moderate increase in the content of azadirachtin $(5.71{\pm}0.34\;mg/L)$ when compared to control cultures $(2.40{\pm}0.56\;mg/L)$. The addition of methyl jasmonate $(500\;{\mu}M/L)$ on day 3 has shown $\~4$ fold improvement in bioproduction of azadirachtln $(6.92{\pm}0.11\;mg/L)$ when compared to control cultures $(1.63{\pm}0.02\;mg/L)$. There was no significant effect of the studied growth retardants and permeabilizing agents on bioproduction of azadirachtin. Cells are cultivated in large volumes using the effective precursors.

• KSBB Journal
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• 제15권4호
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• pp.337-341
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• 2000

The decanter is very useful to harvest biomass from plant cell cultures in large-scale process. It is very important to obtain high yield and low moisture content in recovered biomass so as to minimize solvent usage in subsequent extraction steps. Effluent clarity was also affected by the differential speed although this affect was more dramatic at higher flow rates than at lower flow rates. Moisure content was largely unaffected by flow rate. A decrease in moisture content was evident as differential speed decreased.

• Journal of Plant Biotechnology
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• 제6권2호
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• pp.125-130
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• 2004

Callus and cell suspension cultures of Eurycoma longifolia Jack could be an alternative supply of 9-hydroxycanthin-6-one and 9-methoxycanthin-6-one. The callus tissues were initiated from leaves of different trees. The friable calli were used for the preparation of the cell suspension cultures of E. longifolia. The leaf explant of tree Eu-9 produced the most callus and also induced high cell biomass in the cell suspension culture, but it produced low quantity of 9-methoxycanthin- 6-one and 9-hydroxycanthin-6-one. The leaf explant from tree Eu-8 produced low quantity of callus and cell biomass, but produced the highest quantity of 9-methoxycanthin- 6-one and 9-hydroxycanthin-6-one. Optimum production of cell biomass was obtained on cell culture medium with pH 5.75 prior to autoclaving, but high alkaloids content could be induced in culture medium in acidic condition with pH 4.75 and 5.25 prior to autoclaving.

• Plant Biotechnology Reports
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• 제4권1호
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• pp.9-13
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• 2010

Guggulsterone, a hypolipidemic natural agent, is produced in resin canals of the plant Commiphora wightii. In this study, the stimulatory effects of growth retardants [ALAR (N,N-dimethylaminosuccinamic acid) and CCC (chlormequat chloride)] and fungal elicitor on guggulsterone accumulation in cell cultures of C. wightii are reported. CCC at $1\;mg\;l^{-1}$ enhanced guggulsterone content (${\sim}123\;{\mu}g\;l^{-1})$ when added on the fifth day after inoculation, while ALAR at $2.5\;{\mu}g\;l^{-1}$ increased guggulsterone content (${\sim}116\;{\mu}g\;l^{-1}$) when added on the tenth day. In a two-stage fed-batch process, combined treatment with fungal elicitor and growth retardant caused a significant increase (${\sim}353\;{\mu}g\;l^{-1}$) in guggulsterone content in cell cultures after 17 days of growth. This represents an approximately fivefold increase over the guggulsterone contents in initial cultures of this plant.

• Journal of Plant Biotechnology
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• 제4권4호
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• pp.143-150
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• 2002

A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권3호
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• pp.138-149
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• 2002

Plant cell culture provides a viable alternative over whole plant cultivation for the production of secondary metabolites. In order to successfully cultivate the plant cells at large scale, several engineering parameters such as, cell aggregation, mixing, aeration, and shear sensitivity are taken into account for selection of a suitable bioreactor. The media ingredients, their concentrations and the environmental factors are optimized for maximal synthesis of a desired metabolite. Increased productivity in a bioreactor can be achieved by selection of a proper cultivation strategy (batch, fed-batch, two-stage etc.), feeding of metabolic precursors and extraction of intracellular metabolites. Proper understanding and rigorous analysis of these parameters would pave the way towards the successful commercialization of plant cell bioprocesses.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 춘계학술대회 및 국제심포지움 초록집
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• pp.139-139
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• 2005

• KSBB Journal
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• 제15권4호
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• pp.398-401
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• 2000

Taxus chinensis 식물세포 배양에 의한 paclitaxel대량 생산의 경우 biomass내 paclitaxel이외에 다른 많은 유도체들이 확 인 되었다. 이를 목표 물질로 선정하여 Prep-LC를 이용하여 분리/정제하고 NMR, MS 등으로 화학구조분석을 하여 이들 물질들에 대한 동정을 하였다. 또한 Taxus chinensis 식물세포 배양에 의한 paclitaxel 대량 생산의 경우 paclitaxel 생산량에 따른 여러가지 유도체들의 생산 경향을 조사하였으며, paclitaxel의 생산량이 증가할 수록 유도체들의 생산량은 상대 적으로 감소함을 알 수 있었다. 이러한 연구는 결국 paclitaxel 이외의 새로운 유용성분 및 전구체 확인, 분리/정제 방법의 개발, paclitaxel 생산에서의 품질관리에 유용하게 사용되어 질 수 있다.

• Journal of Microbiology and Biotechnology
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• 제19권6호
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• pp.573-581
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• 2009

The complex enzyme pool secreted by the phytopathogenic fungus Fusarium graminearum in response to glucose or hop cell wall material as sole carbon sources was analyzed. The biochemical characterization of the enzymes present in the supernatant of fungal cultures in the glucose medium revealed only 5 different glycosyl hydrolase activities; by contrast, when analyzing cultures in the cell wall medium, 17 different activities were detected. This dramatic increase reflects the adaptation of the fungus by the synthesis of enzymes targeting all layers of the cell wall. When the enzymes secreted in the presence of plant cell wall were used to hydrolyze pretreated crude plant material, high levels of monosaccharides were measured with yields approaching 50% of total sugars released by an acid hydrolysis process. This report is the first biochemical characterization of numerous cellulases, hemicellulases, and pectinases secreted by F. graminearum and demonstrates the usefulness of the described protein cocktail for efficient enzymatic degradation of plant cell wall.

• 식물조직배양학회지
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• 제25권6호
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• pp.501-505
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• 1998

오이의 국내 F1 품종인 조생낙합의 하배축 유래 배발생 현탁배양 세포의 초저온 보존 시스템을 개발하였다. 액체질소 저장 후 캘러스 재생률은 2M DMNSO와 0.4 M sucrose를 혼용 처리하였을 때 캘러스 재생률이 85%로 가장 높았다. 그러나 glycerol 처리구에서는 처리농도에 상관없이 모든 처리구에서 캘러스 재생이 이루어지지 않았다. 또한 고농도의 삼투용액에서 배양세포의 전처리 과정은 필요하지 않았다. 재생된 캘러스를 1 mg/L 2,4-D가 첨가된 MS 배지로 이식하여 배양하였을 때 다수의 체세포배가 발달하였으며, 체세포배를 MS 기본배지로 옮겨 명배양한 결과 다수의 소식물체가 발달하였다.