Gene expression analysis of enzymes of the carotenoid biosynthesis pathway involved in ß-cryptoxanthin accumulation in wild raspberry, Rubus palmatus.

ß-cryptoxanthin (ß-Cry), a xanthophyll, is unlike other abundant carotenoids, such as α-carotene, ß-carotene, lycopene, lutein, and zeaxanthin. It is not found in most fruits or vegetables but is found only in specific fruits, such as hot chili pepper, persimmon, and citrus fruits. Because recent reports suggest that ß-Cry intake is beneficial to human health, the xanthophyll requires further investigation. Although ß-Cry accumulates in the fruit of wild raspberry, Rubus palmatus, it is not present in cultivated raspberry. In the present study, two wild raspberry species were studied-R. palmatus, which accumulates ß-Cry in the fruit, and R. crataegifolius, which does not accumulate ß-Cry. Four carotenoid biosynthetic enzymes derived from these two species were analyzed-phytoene synthase (PSY), lycopene ß-cyclase (LCYb), ß-carotene hydroxylase (HYb), and zeaxanthin epoxidase (ZEP). Expression levels of their genes were also assessed to elucidate mechanism underlying ß-Cry accumulation. Partial gene sequences of RubPSY, RubLCYb, RubHYb, and RubZEP, isolated from immature raspberry fruits of R. palmatus, were used as probes for Northern blot analysis. RubZEP expression ceased as the fruits matured, possibly because of reduced production of zeaxanthin. ß-Cry is considered to be an intermediate compound that accumulates in the mature fruits of R. palmatus. High expression of RubPSY was detectable in the mature fruits of R. crataegifolius, and the expression of RubLCYb, RubHYb, and RubZEP was detectable during all stages of fruit maturation. In contrast, ß-Cry was absent in the mature fruits of R. crataegifolius.

Modelling the growth kinetics of Kocuria marina DAGII as a function of single and binary substrate during batch production of ß-Cryptoxanthin.

In the present investigation, growth kinetics of Kocuria marina DAGII during batch production of ß-Cryptoxanthin (ß-CRX) was studied by considering the effect of glucose and maltose as a single and binary substrate. The importance of mixed substrate over single substrate has been emphasised in the present study. Different mathematical models namely, the Logistic model for cell growth, the Logistic mass balance equation for substrate consumption and the Luedeking-Piret model for ß-CRX production were successfully implemented. Model-based analyses for the single substrate experiments suggested that the concentrations of glucose and maltose higher than 7.5 and 10.0 g/L, respectively, inhibited the growth and ß-CRX production by K. marina DAGII. The Han and Levenspiel model and the Luong product inhibition model accurately described the cell growth in glucose and maltose substrate systems with a R 2 value of 0.9989 and 0.9998, respectively. The effect of glucose and maltose as binary substrate was further investigated. The binary substrate kinetics was well described using the sum-kinetics with interaction parameters model. The results of production kinetics revealed that the presence of binary substrate in the cultivation medium increased the biomass and ß-CRX yield significantly. This study is a first time detailed investigation on kinetic behaviours of K. marina DAGII during ß-CRX production. The parameters obtained in the study might be helpful for developing strategies for commercial production of ß-CRX by K. marina DAGII.