195
1989 – unfortunately – the continuous production of cyclodextrins by using immobilized enzyme has not been scaled up on process scale.
4.2.2 Production of Beta-Cyclodextrin at Chinoin
Laboratory processing: The laboratory-scale experiments for development of betacyclodextrin production technology started in about 1977. The fundamentals of the
technology have not changed since Schardinger (1903): A cyclodextrin glucosyltransferase enzyme of B. macerans origin was used. It was clear, at the first occasion, that foamy, stinking, and not easy-to-handle culture liquors of B. macerans
were not suitable for an economical high-throughput industrial manufacturing.
After optimizing the fermentation, crude enzyme filtering and fractionating parameters of the downstream processing were established, and a robust and reproducible
enzyme technology was documented in 1982. Compared to achievable 8.7% and
10.2% cyclodextrin yields using crude enzyme powder supply, conversion rates of
beta-cyclodextrin production increased to 12.6% and 17.5% after 3- and 5-day incubation, respectively, with a semi-purified and hydrolase-free enzyme preparation
(Seres et al. 1980).
Beta-cyclodextrin manufacturing technology: In the early 1980s, only Japanese
firms distributed limited quantities of purified beta-cyclodextrin in the world market: (i) Celdex-N from Nihon Shokuhin Kako (a company belonging to Mitsubishi,
Japan); (ii) Ringdex from Ocean Sanraku (a company belonging to Sumitomo,
Japan); and (iii) Dexypearl from Ensuiko Sugar Refining Co. Ltd. Japan.
The initial concept was based on the common accepted fact that continuous precipitation of water-insoluble toluene/beta-cyclodextrin complex shifted the conversion direction toward beta-cyclodextrin production. After preliminary laboratory
procedures using potato starch, corn starch was selected as raw substrate because of
the higher abundance in Hungary and its lower price. Due to its shorter chain and
more branched structure of corn amylopectin, reduced viscosities could be achieved
after gelation of the high-starch-containing gel. This gelatinization process makes
possible to prepare a concentrated (20–45 w/w %) gelatinized starch solution with
considerable viscosities without any observable retrogradation of starch and provides high cyclodextrin levels in the further cyclodextrin glucosyltransferasecatalyzed conversion (Vakaliu et al. 1979). An optimal starch concentration of
around 30% was found to give the highest cyclodextrin yield. Additional step was
inserted into the process to hydrolyze the resulted starch gel to 1–3 dextrose equivalents. For the sake of this mild hydrolysis, an alpha-amylase of Bacillus subtilis
origin was applied. Existing calcium ions are essential to stimulate – as cofactor –
both alpha-amylase and cyclodextrin glucosyltransferase activity. The optimal pH
(7.2) and the essential cofactor input were maintained with sodium hydroxide and
calcium carbonate. The cyclodextrin glucosyltransferase used in this process was
the own product of Chinoin.
4 History of Cyclodextrin Production in Hungary
1989 – unfortunately – the continuous production of cyclodextrins by using immobilized enzyme has not been scaled up on process scale.
4.2.2 Production of Beta-Cyclodextrin at Chinoin
Laboratory processing: The laboratory-scale experiments for development of betacyclodextrin production technology started in about 1977. The fundamentals of the
technology have not changed since Schardinger (1903): A cyclodextrin glucosyltransferase enzyme of B. macerans origin was used. It was clear, at the first occasion, that foamy, stinking, and not easy-to-handle culture liquors of B. macerans
were not suitable for an economical high-throughput industrial manufacturing.
After optimizing the fermentation, crude enzyme filtering and fractionating parameters of the downstream processing were established, and a robust and reproducible
enzyme technology was documented in 1982. Compared to achievable 8.7% and
10.2% cyclodextrin yields using crude enzyme powder supply, conversion rates of
beta-cyclodextrin production increased to 12.6% and 17.5% after 3- and 5-day incubation, respectively, with a semi-purified and hydrolase-free enzyme preparation
(Seres et al. 1980).
Beta-cyclodextrin manufacturing technology: In the early 1980s, only Japanese
firms distributed limited quantities of purified beta-cyclodextrin in the world market: (i) Celdex-N from Nihon Shokuhin Kako (a company belonging to Mitsubishi,
Japan); (ii) Ringdex from Ocean Sanraku (a company belonging to Sumitomo,
Japan); and (iii) Dexypearl from Ensuiko Sugar Refining Co. Ltd. Japan.
The initial concept was based on the common accepted fact that continuous precipitation of water-insoluble toluene/beta-cyclodextrin complex shifted the conversion direction toward beta-cyclodextrin production. After preliminary laboratory
procedures using potato starch, corn starch was selected as raw substrate because of
the higher abundance in Hungary and its lower price. Due to its shorter chain and
more branched structure of corn amylopectin, reduced viscosities could be achieved
after gelation of the high-starch-containing gel. This gelatinization process makes
possible to prepare a concentrated (20–45 w/w %) gelatinized starch solution with
considerable viscosities without any observable retrogradation of starch and provides high cyclodextrin levels in the further cyclodextrin glucosyltransferasecatalyzed conversion (Vakaliu et al. 1979). An optimal starch concentration of
around 30% was found to give the highest cyclodextrin yield. Additional step was
inserted into the process to hydrolyze the resulted starch gel to 1–3 dextrose equivalents. For the sake of this mild hydrolysis, an alpha-amylase of Bacillus subtilis
origin was applied. Existing calcium ions are essential to stimulate – as cofactor –
both alpha-amylase and cyclodextrin glucosyltransferase activity. The optimal pH
(7.2) and the essential cofactor input were maintained with sodium hydroxide and
calcium carbonate. The cyclodextrin glucosyltransferase used in this process was
the own product of Chinoin.
4 History of Cyclodextrin Production in Hungary