PAOX1 expression in mixed-substrate continuous cultures of Komagataella phaffii (Pichia pastoris) is completely determined by methanol uptake regardless of the secondary carbon source: a simplifying principle for predicting recombinant protein production.

The expression of recombinant proteins by the AOX1 promoter of Komagataella phaffii is typically induced by adding methanol to the cultivation medium. Since growth on methanol imposes a high oxygen demand, the medium is often supplemented with an additional "secondary" carbon source which serves to reduce the consumption of methanol, and hence, oxygen. Early research recommended the use of glycerol as the secondary carbon source, but more recent studies recommend the use of sorbitol because glycerol represses PAOX1 expression. To assess the validity of this recommendation, we measured the steady state concentrations of biomass, residual methanol, and AOX1 over a wide range of dilution rates (0.02-0.20 h-1) in continuous cultures of the Mut+ strain fed with methanol, methanol + glycerol, and methanol + sorbitol. We find that when the specific AOX1 expression and methanol uptake rates for each of the three feeds are plotted against each other, they collapse into a single hyperbolic curve. The specific AOX1 expression rate is therefore completely determined by the specific methanol uptake rate regardless of the existence (present/absent) and type (repressing/non-repressing) of the secondary carbon source. In particular, cultures fed with methanol + glycerol and methanol + sorbitol that consume methanol at equal rates also express the protein at equal rates and levels. Now, it turns out that the simple unstructured model developed by Egli and co-workers can predict the specific methanol uptake rates of single- and mixed-substrate cultures over a wide range of dilution rates and feed concentrations. By combining this model with our data, we derive simple formulas that predicts the protein expression rates and levels of single- and mixed-substrate cultures over a wide range of conditions.

Co-metabolic biodegradation of 4-bromophenol in mixture of pollutants system by Arthrobacter chlorophenolicus A6

Brominated phenols are listed as priority pollutants, and are the key components of paper pulp wastewater together with nitrophenol and chlorophenol. However, the biodegradation of bromophenol in a mixed substrate system is very scanty. In the present investigation, simultaneous biodegradation kinetics of three substituted phenols (4-bromophenol, 4-BP; 4-nitrophenol, 4-NP; and 4-chlorophenol, 4-CP) were investigated using Arthrobacter chlorophenolicus A6. A 23 full factorial design was applied with varying 4-BP and 4-CP from 75–125 mgl− 1and4-NP from 50–100 mgl− 1. Almost complete degradation of this mixture of substituted phenols was achieved at an initial concentration combination of 125, 125, and 100 mgl− 1of 4-CP, 4-BP, and 4-NP, respectively in 68 h. Statistical analysis of the results revealed that among the three variables, 4-NP had the most prominent influence on both degradations of 4-CP and 4-BP. While the concentration of 4-CP had a strong negative interaction effect on the biodegradation of 4-NP. Irrespective of the concentration levels of these three substrates, 4-NP was preferentially biodegraded over 4-CP and 4-BP. Further, 4-BP biodegradation rates were found to be higher than that of 4-CP followed by 4-NP. Besides, the variation of biomass yield coefficient of the culture was investigated at different initial concentration combinations of these substituted phenols. Although the actinomycetes consumed 4-NP at a faster rate, the biomass yield was very poor. This revealed that the microbial cells were more stressed when grown on 4-NP compared to 4-BP and 4-CP. Overall, this study revealed the prospective of A. chlorophenolicus A6 for the degradation of 4-BP in mixed substrate systems.

Influence of Mixed Substrate and Arbuscular Mycorrhizal Fungi on Photosynthetic Efficiency, Nutrient and Water Status and Yield in Tomato Plants Irrigated with Saline Reclaimed Waters

The use of reclaimed water (RW) is considered as a means of maintaining agricultural productivity under drought conditions. However, RW may contain high concentrations of salts. The use of some practices, such as biofertilizers and organic substrates, is also becoming increasingly important in agricultural. production. The aim of this study was to evaluate the application of a mixed substrate (with coconut fibre) and arbuscular mycorrhizal fungi (AMF) on water relations, nutrient uptake and productivity in tomato plants irrigated with saline RW in a commercial greenhouse. Saline RW on its own caused a nutrient imbalance and negatively affected several physiological parameters. However, the high water-holding capacity of coconut fibre in the mixed substrate increased water and nutrient availability for the plants. As a consequence, leaf water potential, gas exchange, some fluorescence parameters (PhiPSII, Fv’/Fm’, qP and ETR) and fruit size and weight improved, even in control irrigation conditions. The use of AMF improved only some parameters because of the low percentage of colonization, suggesting that AMF effectiveness in commercial field conditions is slower and dependent of several factors.