DESIGN: Food choice was assessed using the validated New Zealand Adolescent FFQ. Principal components analysis was used to determine dietary patterns. Trained research assistants measured participants' height and body mass. Cardiorespiratory fitness was assessed in a subset of participants using the multistage 20 m shuttle run. The level and stage were recorded, and the corresponding VO2max was calculated. Differences in mean VO2max according to sex and BMI were assessed using t tests, while associations between cardiorespiratory fitness and dietary patterns were examined using linear regression analyses adjusted for age, sex, school attended, socio-economic deprivation and BMI.
SETTING: Secondary schools in Otago, New Zealand.
SUBJECTS: Students (n 279) aged 14-18 years who completed an online lifestyle survey during a class period.
RESULTS: Principal components analysis produced three dietary patterns: 'Treat Foods', 'Fruits and Vegetables' and 'Basic Foods'. The 279 participants who provided questionnaire data and completed cardiorespiratory fitness testing had a mean age of 15·7 (sd 0·9) years. Mean VO2max was 45·8 (sd 6·9) ml/kg per min. The 'Fruits and Vegetables' pattern was positively associated with VO2max in the total sample (β=0·04; 95%CI 0·02, 0·07), girls (β=0·06; 95% CI 0·03, 0·10) and boys (β=0·03; 95% CI 0·01, 0·05).
CONCLUSIONS: These results indicate that increase in cardiorespiratory fitness was associated with a healthier dietary pattern, suggesting both should be targeted as part of a global lifestyle approach. Longitudinal studies are needed to confirm this association in relation to health outcomes in New Zealand adolescents.
METHODS: In this study, the gene encoding a cellobiohydrolase B (cbhB) from A. niger ATCC 10574 was cloned and expressed in the methylotrophic yeast Pichia pastoris X-33. The recombinant CBHB was purified and characterised to study its biochemical and kinetic characteristics. To evaluate the potential of CBHB in assisting biomass conversion, CBHB was supplemented into a commercial cellulase preparation (Cellic(®) CTec2) and was used to hydrolyse oil palm empty fruit bunch (OPEFB), one of the most abundant lignocellulosic waste from the palm oil industry. To attain maximum saccharification, enzyme loadings were optimised by response surface methodology and the optimum point was validated experimentally. Hydrolysed OPEFB samples were analysed using attenuated total reflectance FTIR spectroscopy (ATR-FTIR) to screen for any compositional changes upon enzymatic treatment.
RESULTS: Recombinant CBHB was over-expressed as a hyperglycosylated protein attached to N-glycans. CBHB was enzymatically active towards soluble substrates such as 4-methylumbelliferyl-β-D-cellobioside (MUC), p-nitrophenyl-cellobioside (pNPC) and p-nitrophenyl-cellobiotrioside (pNPG3) but was not active towards crystalline substrates like Avicel(®) and Sigmacell cellulose. Characterisation of purified CBHB using MUC as the model substrate revealed that optimum catalysis occurred at 50 °C and pH 4 but the enzyme was stable between pH 3 to 10 and 30 to 80 °C. Although CBHB on its own was unable to digest crystalline substrates, supplementation of CBHB (0.37%) with Cellic(®) CTec2 (30%) increased saccharification of OPEFB by 27%. Compositional analyses of the treated OPEFB samples revealed that CBHB supplementation reduced peak intensities of both crystalline cellulose Iα and Iβ in the treated OPEFB samples.
DISCUSSION: Since CBHB alone was inactive against crystalline cellulose, these data suggested that it might work synergistically with other components of Cellic(®) CTec2. CBHB supplements were desirable as they further increased hydrolysis of OPEFB when the performance of Cellic(®) CTec2 was theoretically capped at an enzyme loading of 34% in this study. Hence, A. niger CBHB was identified as a potential supplementary enzyme for the enzymatic hydrolysis of OPEFB.