High-level phenol bioproduction by engineered Pichia pastoris in glycerol fed-batch fermentation using an efficient pertraction system.

This study aimed to establish a high-level phenol bioproduction system from glycerol through metabolic engineering of the yeast Pichia pastoris (Komagataella phaffii). Introducing tyrosine phenol-lyase to P. pastoris led to a production of 59 mg/L of phenol in flask culture. By employing a strain of P. pastoris that overproduces tyrosine-a precursor to phenol-we achieved a phenol production of 1052 mg/L in glycerol fed-batch fermentation. However, phenol concentrations exceeding 1000 mg/L inhibited P. pastoris growth. A phenol pertraction system utilizing a hollow fiber membrane contactor and tributyrin as the organic solvent was developed to reduce phenol concentration in the culture medium. Integrating this system with glycerol fed-batch fermentation resulted in a 214 % increase in phenol titer (3304 mg/L) compared to glycerol fed-batch fermentation alone. These approaches offer a significant framework for the microbial production of chemicals and materials that are highly toxic to microorganisms.

Preparation of Chemically Resistant Cellulose Benzoate Hollow Fiber Membrane via Thermally Induced Phase Separation Method.

For the first time, we have successfully fabricated microfiltration (MF) hollow fiber membranes by the thermally induced phase separation (TIPS) and non-solvent induced phase separation (NIPS) methods using cellulose acetate benzoate (CBzOH), which is a cellulose derivative with considerable chemical resistance. To obtain an appropriate CBzOH TIPS membrane, a comprehensive solvent screening was performed to choose the appropriate solvent to obtain a membrane with a porous structure. In parallel, the CBzOH membrane was prepared by the NIPS method to compare and evaluate the effect of membrane structure using the same polymer material. Prepared CBzOH membrane by TIPS method showed high porosity, pore size around 100 nm or larger and high pure water permeability (PWP) with slightly low rection performance compared to that by NIPS. On the contrary, CBzOH membranes prepared with the NIPS method showed three times lower PWP with higher rejection. The chemical resistance of the prepared CBzOH membranes was compared with that of cellulose triacetate (CTA) hollow fiber membrane, which is a typical cellulose derivative as a control membrane, using a 2000 ppm sodium hypochlorite (NaClO) solution. CBzOH membranes prepared with TIPS and NIPS methods showed considerable resistance against the NaClO solution regardless of the membrane structure, porosity and pore size. On the other hand, when the CTA membrane, as the control membrane, was subjected to the NaClO solution, membrane mechanical strength sharply decreased over the exposure time to NaClO. It is interesting that although the CBzOH TIPS membrane showed three times higher pure water permeability than other membranes with slightly lower rejection and considerably higher NaClO resistance, the mechanical strength of this membrane is more than two times higher than other membranes. While CBzOH samples showed no change in chemical structure and contact angle, CTA showed considerable change in chemical structure and a sharp decrease in contact angle after treatment with NaClO. Thus, CBzOH TIPS hollow fiber membrane is noticeably interesting considering membrane performance in terms of filtration performance, mechanical strength and chemical resistance on the cost of slightly losing rejection performance.