Since the mid-1960s, methylotrophic yeast Komagataella phaffii (previously described as Pichia pastoris) has received increasing scientific attention. The interest for the industrial production of proteins for different applications (e.g., feed, food additives, detergent, waste treatment processes, and textile) is a well-consolidated scientific topic, and the importance for this approach is rising in the current era of environmental transition in human societies. This review aims to summarize fundamental and specific information in this scientific field. Additionally, an updated description of the relevant products produced with K. phaffii at industrial levels by a variety of companies—describing how the industry has leveraged its key features, from products for the ingredients of meat-free burgers (e.g., IMPOSSIBLE™ FOODS, USA) to diabetes therapeutics (e.g., Biocon, India)—is provided. Furthermore, active patents and the typical workflow for industrial protein production with this strain are reported.
The yeast Yarrowia lipolytica is an industrially relevant microorganism, which is able to convert low-value wastes into different high-value, bio-based products, such as enzymes, lipids, and other important metabolites. Waste cooking oil (WCO) represents one of the main streams generated in the food supply chain, especially from the domestic sector. The need to avoid its incorrect disposal makes this waste a resource for developing bioprocesses in the perspective of a circular bioeconomy. To this end, the strain Y. lipolytica W29 was used as a platform for the simultaneous production of intracellular lipids and extracellular lipases. Three different minimal media conditions with different pH controls were utilized in a small-scale (50 mL final volume) screening strategy, and the best condition was tested for an up-scaling procedure in higher volumes (800 mL) by selecting the best-performing possibility. The tested media were constituted by YNB media with high nitrogen restriction (1 g L−1 (NH4)2SO4) and different carbon sources (3% w v−1 glucose and 10% v v−1 WCO) with different levels of pH controls. Lipase production and SCO content were analyzed. A direct correlation was found between decreasing FFA availability in the media and increasing SCO levels and lipase activity. The simultaneous production of extracellular lipase (1.164 ± 0.025 U mL−1) and intracellular single-cell oil accumulation by Y. lipolytica W29 growing on WCO demonstrates the potential and the industrial relevance of this biorefinery model.
The edible plant oils production is associated with the release of different types of by-products. The latter represent cheap and available substrates to produce valuable compounds, such as flavours and fragrances, biologically active compounds and bio-based polymers. Elizabethkingia meningoseptica Oleate hydratases (Em_OhyA) can selectively catalyze the conversion of unsaturated fatty acids, specifically oleic acid, into hydroxy fatty acids, which find different industrial applications. In this study, Design-of-experiment (DoE) strategy was used to screen and identify conditions for reaching high yields in the reaction carried out by Escherichia coli whole-cell carrying the recombinant enzyme Em_OhyA using Waste Cooking Oils (WCO)-derived free fatty acids (FFA) as substrate. The identified reaction conditions for high oleic acid conversion were also tested on untreated triglycerides-containing substrates, such as pomace oil, sunflower oil, olive oil and oil mill wastewater (OMW), combining the triglyceride hydrolysis by the lipase from Candida rugosa and the E. coli whole-cell containing Em_OhyA for the production of hydroxy fatty acids. When WCO, sunflower oil and OMW were used as substrate, the one-pot bioconversion led to an increase of oleic acid conversion compared to the standard reaction. This work highlights the efficiency of the DoE approach to screen and identify conditions for an enzymatic reaction for the production of industrially-relevant products.