• KSBB Journal
• /
• v.8 no.5
• /
• pp.488-494
• /
• 1993

The accumulation of benzophenanthridine alkaloids, sanguinarine, chelerythrine, chelirubine and macarpine occurred in suspension cultures of Eschscholtzia californica. To increase alkaloid production, feeding experiments with the biosynthetic precursors, tyrosine, tyramine, L-dopa, dopamine with and without elicitation were studied. In feeding experiments with various precursors, the total alkaloid production was slightly increased. The precursor feeding with elicitation, however, increased total alkaloid production several times.

• KSBB Journal
• /
• v.13 no.6
• /
• pp.700-705
• /
• 1998

Studies were made to investigate effects of the scale-up of stirred tank bioreactors on cell growth and alkaloids production for suspension cultures of Eschscholtzia californica. In the 1.5 L STR, cell lysis was observed at 110 rpm or higher agitation speed. The agitation speed of 30 L STR was 43.7 rpm to maintain the same shear stress developed in 1.5 L STR of 100 rpm. As a result of scale-up from 1.5 L to 30 L STR, the specific growth rate was decreased from 0.12 to 0.07 day-1. The alkaloids productivity was also decreased from 0.24 to 0.14 mg/L-day. Changes of mixing performance and oxygen transfer were studied to explain the decrease of cell growth and alkaloids production. Decreased oxygen transfer rate coefficient(KLa) and increased mixing time by the scale-up was observed at various aeration rates.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.1
• /
• pp.107-110
• /
• 2000

Elicited cells with methyl jasmonate continued to produce benzophenanthridine alkaloids throughout medium changes in suspension cultures of Eschscholtzia californica. Large increases in alkaloid production were observed by re-elitations with medium changes. The total alkaloid production increased during the successive elicitation steps reaching a maximum level on the 4th elicitation. The highest total alkaloid produced was 250 mg/I, which was 20fold higher than that of the single elicitation and 4-fold higher than that of the normal culture without elicitation. The large increases in alkaloid production in successive re-elicitations with medium changes are believed to be caused by the accumulation of the signal transduction compound, jasmonate.

• Journal of Microbiology and Biotechnology
• /
• v.3 no.4
• /
• pp.238-243
• /
• 1993

In the suspension cultures of Eschscholtzia californica, phytohormone effects showed that alkaloid production was increased by IAA treatment without kinetin in both volumetric and specific way. Kinetin, however, suppressed alkaloid accumulation. Addition of ethephon inhibited cell growth. However, it enhanced the alkaloid production significantly in both volumetric and specific way. IAA promoted alkaloid production during elicitation. The highest alkaloid accumulation was observed at 5 $\mu$ M of IAA. Ethephon also enhanced alkaloid production during elicitation. The highest alkaloid formation was observed at 460 mg/l of ethephon with elicitation. Elicitation with ethephon, however, altered cell growth and the pattern of benzophenanthridine alkaloids production.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.2
• /
• pp.255-262
• /
• 2008

Methyl jasmonate (MJ) and yeast extract (YE) induce protein expression and benzophenanthridine alkaloid accumulation in Eschscholtzia californica suspension cell cultures. One hundred ${\mu}M$ MJ primarily induced dihydrosanguinarine $(509.0{\pm}7.4mg/l)$ ; 0.2g/l YE induced sanguinarine $(146.8{\pm}3.8mg/l)$ and an unknown compound. These results occur because dihydrobenzophenanthridine oxidase (DHBO) is induced by YE and not by MJ. YE and chitin (CHI) had similar effects on sanguinarine production and DHBO expression. Differential induction of secondary metabolites was shown in E. californica suspension cultures and the expression of proteins confirmed the metabolite results. Furthermore, treatment by various oligosaccharides helped us to understand the elicitation effect of YE in signal transduction pathways.

• KSBB Journal
• /
• v.9 no.2
• /
• pp.180-185
• /
• 1994

The productivity of alkaloid in the airlift fermentor operation was less than that of suspension coltures of Eschscholtzia californica cells in the shake flask. To overcome the productivity reduction, a gas recycle airlift fermentor was developed because the gas-stripping in normal airlift fermentor was believed to play a significant role for productivity reduction. The alkaloid content in the gas recycle system with Eschscholtzia californica suspension cells was 2.7 times higher than that of normal airlift fermentor. The productivity of alkaloids and $CO_2$ concentration were affected by the volume of gas reservoir in the gas recycle airlift fermentor.

• Journal of Microbiology and Biotechnology
• /
• v.1 no.3
• /
• pp.220-226
• /
• 1991

Large and rapid increases in benzophenanthridine alkaloid production occured in suspension cultures of Eschscholtzia californica cells treated with elicitors. Response to different biotic elicitors showed that elicitors prepared from yeast extract, Collectotrichum lindemuthianum and Verticillium dahliae induced alkaloid formation. Highest alkaloid accumulation was obtained with $60\;\mu\textrm{g}$ of yeast extract elicitor per gram of fresh cell weight. In time course performance after elicitor addition, more than 40 hours were required to obtain saturated alkaloid accumulation. Compounded silicone fluid, an ideal accumulation phase for two-phase culture of E. californica, accumulated a large amount of alkaloids produced in a specific manner. Elicitation in two-phase culture clearly increased net alkaloid production as well as their concentrations in the accumulation phase.

• KSBB Journal
• /
• v.24 no.2
• /
• pp.195-200
• /
• 2009

The effect of nitrogen source on cell growth and benzo[c]phenanthridine alkaloids production by modifying $NO_3\;^-:NH_4\;^+$ ratio in cell suspension culture of Eschscholtzia califarnica was investigated. When total nitrogen concentration is maintained (60 mM), maximum benzo[c]phenanthridine alkaloids production is about 60.72 mg/L at 50:10 (mol/mol). This productivity was 3.8 times higher than that obtained when cells were grown instandard MS medium. The decrease of $NO_3\;^-:NH_4\;^+$ ratio at 60 mM of total nitrogen caused the decline of both growth and benzo[c]phenanthridine alkaloids production. Under the same concentration of $N0_3\;^-$ (50 mM), higher concentration of $NH_4\;^+$ inhibited cell growth strongly but induced alkaloids production slightly. Also, under the same concentration of $NH_4\;^+$ (25 mM), higher concentration of $N0_3\;^-$ induced alkaloids production strongly but high concentration of $N0_3\;^-$ (${\geq}$100 mM) interfered alkaloids instead. Maximum benzo[c]phenanthridine alkaloids production is about 62.71 mg/L at 50:25 (mol/mol). These results suggest that higher biomass and higher alkaloids production could be obtained by optimizing each nitrogen concentration as well as $NO_3\;^-:NH_4\;^+$ ratio in the culture medium. Nitrate and ammonium in culture medium have distinct role in the regulation of growth and alkaloids production; ammonium had a strong influence on growth while nitrate had an influence on alkaloids production.

• KSBB Journal
• /
• v.20 no.4
• /
• pp.285-290
• /
• 2005

Benzophenanthridine alkaloids - sanguinarine, chelirubine, macarpine, and chelerythrine are produced from Eschscholtzia californica (Californica Poppy, used as a sedative by Native Americans) and most of them are derived from dihydrosanguinarine. The properties of sanguinarine are the basis of its antimicrobial activity and its use in chemosurgery and skin cancer excision. For overproduction of sanguinarine from E. californica, yeast extract was used as elicitor and the elicited cell's metabolites were checked. Sanguinarine production was increased intracelluarly about 8 times in the cell and 5 times extracelluarly. We have peformed proteomic analysis of proteins sequentially extracted from E. califormica suspended cells which were cultured with elicitor, an increase of spot intensity was seen at 24 hours following elicitation. These proteins were separated by two-dimensional electrophoresis (2-DE). We found several spots that were expected to be related to benzophenanthridine alkaloids production by comparing the production profiles of metabolites such as sanguinarine. These results demonstrate the use of metabolite analysis as a tool for detecting target proteins related to metabolites production pathway.

• 한국생물공학회:학술대회논문집
• /
• 2003.04a
• /
• pp.103-104
• /
• 2003