Meat analogue research and development focuses on the production of sustainable products that recreate conventional meat in its physical sensations (texture, appearance, taste, etc.) and nutritional aspects. Minced products, like burger patties and nuggets, muscle-type products, like chicken or steak-like cuts, and emulsion products, like Frankfurter and Mortadella type sausages, are the major categories of meat analogues. In this review, we discuss key ingredients for the production of these novel products, with special focus on protein sources, and underline the importance of ingredient functionality. Our observation is that structuring processes are optimized based on ingredients that were not originally designed for meat analogues applications. Therefore, mixing and blending different plant materials to obtain superior functionality is for now the common practice. We observed though that an alternative approach towards the use of ingredients such as flours, is gaining more interest. The emphasis, in this case, is on functionality towards use in meat analogues, rather than classical functionality such as purity and solubility. Another trend is the exploration of novel protein sources such as seaweed, algae and proteins produced via fermentation (cellular agriculture).

Functionality of Ingredients and Additives in Plant-Based Meat Analogues

Haem has widespread applications in healthcare and food supplement industries. Escherichia coli has previously been engineered to produce a small amount of haem intracellularly through the C4 pathway, requiring extraction for applications. Here we report secretory production of free haem by engineered E. coli strains, using the C5 pathway and the optimized downstream pathway for haem biosynthesis. Furthermore, knocking out ldhA, pta and also yfeX—encoding a putative haem-degrading enzyme—results in 7.88 mg l−1 of total haem with 1.26 mg l−1 of extracellular haem in flask cultivation. Fed-batch fermentations of the engineered strain overexpressing a haem exporter CcmABC from glucose only and glucose supplemented with l-glutamate secrete 73.4 and 151.4 mg l−1 of haem, respectively, which are 63.5% of 115.5 mg l−1 and 63.3% of 239.2 mg l−1 of total haem produced. The engineered E. coli strain reported here will be useful for microbial production of free haem. Microbial production of haem for applications in healthcare and food supplement industry requires high-performing strains. Here, Lee and co-workers report secretory production of free haem by metabolically engineered Escherichia coli strains to produce up to 239 mg l−1 total haem.

Metabolic engineering of Escherichia coli for secretory production of free haem

The present disclosure provides a HTP microbial genomic engineering platform that is computationally driven and integrates molecular biology, automation, and advanced machine learning protocols. This integrative platform utilizes a suite of HTP molecular tool sets to create HTP genetic design libraries, which are derived from, inter alia, scientific insight and iterative pattern recognition. The HTP genomic engineering platform described herein is microbial strain host agnostic and therefore can be implemented across taxa. Furthermore, the disclosed platform can be implemented to modulate or improve any microbial host parameter of interest.

Microbial strain improvement by a htp genomic engineering platform

The present disclosure describes methods for generating microbial strains expressing a heterologous bacterial hemoglobin gene that produce biomolecules of interest. In aspects, the disclosure provides novel bacterial strains, which express a heterologous bacterial hemoglobin gene whose expression is controlled by a native Corynebacterium glutamicum promoter or a mutant promoter derived therefrom. Also provided herein are methods for producing a library of bacterial hemoglobin genes using a promoter ladder comprising a plurality of promoters derived from Corynebacterium glutamicum.

Methods for generating a bacterial hemoglobin library and uses thereof

The present disclosure describes methods for generating microbial strains expressing a heterologous bacterial glucose permease gene that produce biomolecules of interest. In aspects, the disclosure provides novel bacterial strains, which express a heterologous bacterial glucose permease gene whose expression is controlled by a native Corynebacterium glutamicum promoter or a mutant promoter derived therefrom. Also provided herein are methods for producing a library of bacterial glucose permease genes using a promoter ladder comprising a plurality of promoters derived from Corynebacterium glutamicum .

Methods for generating a glucose permease library and uses thereof

This disclosure provides for methods and compositions for the expression and secretion of heme-containing polypeptides.

Secretion of heme-containing polypeptides

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Secretion of heme-containing polypeptides