Microbial Fermentation Process Research Articles

Production of hydrolysates from unmilled AFEX-pretreated switchgrass and comparative fermentation with Zymomonas mobilis

The biological route for producing biofuel from lignocellulosic biomass involves many steps, including biomass harvesting and processing, pretreatment, enzymatic hydrolysis, microbial fermentation, and other downstream processes. Milling to reduce biomass fragment size is a common processing step, which aids subsequent biomass pretreatment and hydrolysis. However, milling also adds additional time and cost. We report here the production of 7% glucan loading hydrolysates using milled and unmilled Ammonia-Fiber Expansion (AFEX)-pretreated switchgrass (7% ASGH). No significant differences in fermentation performance by Zymomonas mobilis were observed between milled and unmilled switchgrass hydrolysates. We also produced a higher 10.5% glucan loading ASGH (10.5% ASGH) using unmilled pretreated switchgrass, and found that xylose fermentation by Z. mobilis was greatly inhibited in the 10.5% ASGH. The chemical compositions of hydrolysates from unmilled and milled switchgrass were analyzed and compared.

Feature-Based Molecular Networking Analysis of the Metabolites Produced by In Vitro Solid-State Fermentation Reveals Pathways for the Bioconversion of Epigallocatechin Gallate

Dark teas are prepared by a microbial fermentation process. Flavan-3-ol B-ring fission analogues (FBRFAs) are some of the key bioactive constituents that characterize dark teas. The precursors and the synthetic mechanism involved in the formation of FBRFAs are not known. Using a unique solid-state fermentation system with β-cyclodextrin inclusion complexation as well as targeted chromatographic isolation, spectroscopic identification, and Feature-based Molecular Networking on the Global Natural Products Social Molecular Networking web platform, we reveal that dihydromyricetin and the FBRFAs, including teadenol A and fuzhuanin A, are derived from epigallocatechin gallate upon exposure to fungal strains isolated from Fuzhuan brick tea. In particular, the strains from subphylum Pezizomycotina were key drivers for these B-/C-ring oxidation transformations. These are the same transformations seen during the fermentation process of dark teas. These discoveries set the stage to enrich dark teas and other food products for these health-promoting constituents.

English

In the last decade, bioethanol has become a powerful biofuel for the improvement of environmental pollution such as reduction in greenhouse gas levels. Yet, the source and type of substrate material plays a crucial role in the bioethanol production process due to the different compositional characteristics and availability of monomeric sugars. Different substrates of first, second and third generation fuel sources exist and may be used as reliable and sustainable substrates for bioethanol generation. The current review provides an overview of Vitellaria fruit pulp; its composition and characteristics for ethanol production. This study has examined literature on the background of the Vitellaria paradoxa, the characteristics and the potential of the shea nut pulp for fermentation to bioethanol. This review will be useful in harnessing the potentials of the shea pulp as industrially relevant substrate for use independently or in combination with other substrates in microbial fermentation processes for ethanol production.   Key words: Vitellaria, shea nut pulp, fermentation, composition, characterization, bioethanol.

Economic Process Evaluation and Environmental Life-Cycle Assessment of Bio-Aromatics Production.

The bio-based production of aromatics is experiencing a renaissance with systems and synthetic biology approaches promising to deliver bio-catalysts that will reach yields, rates, and titers comparable to already existing bulk bio-processes for the production of amino acids for instance. However, aromatic building blocks derived from petrochemical routes have a huge economic advantage, they are cheap, and very cheap in fact. In this article, we are trying to shed light on an important aspect of biocatalyst development that is frequently overlooked when working on strain development: economic and environmental impact of the production process. We estimate the production cost and environmental impact of a microbial fermentation process depending on culture pH, carbon source and process scale. As a model molecule we use para-hydroxybenzoic acid (pHBA), but the results are readily transferrable to other shikimate derived aromatics with similar carbon yields and production rates.

Production of plant-derived anticancer precursor glucoraphanin in chromosomally engineered Escherichia coli

Glucoraphanin is a methionine-derived glucosinolate that imparts numerous health-benefits with broad bioactivity. Low amounts in plant tissues and high cost of extraction have limited the production of glucoraphanin. Metabolic engineering in heterologous microorganisms is an attractive approach to achieve efficient production of valuable natural products. In this study, a microbial fermentation process for glucoraphanin production was demonstrated. The engineered bacterial strain stably expressed 10 allogeneic enzymes in E. coli chromosome, including nine heterologous genes from Arabidopsis and Brassica and one from fungus Neurospora crassa, which could produce the specialized glucosinolate compound glucoraphanin with a titer of 0.675 μg/L by fermentation from glucose. The cofactor supplements and individual gene overexpression for glucoraphanin production were also investigated. This work highlights the possibility of supplying specialized plant glucosinolates by microbial fermentation process, instead of chemical extraction. Additionally, the limiting step enzyme, UDP-glucose-thiohydroximate glucosyltransferase, identified in this study also laid a foundation for further optimizing the glucoraphanin-producing cell factory.

Identification of an unauthorized genetically modified bacteria in food enzyme through whole-genome sequencing

Recently, the unexpected presence of a viable unauthorized genetically modified bacterium in a commercialized food enzyme (protease) product originating from a microbial fermentation process has been notified at the European level (RASFF 2019.3332). This finding was made possible thanks to the use of the next-generation sequencing technology, as reported in this study. Whole-genome sequencing was used to characterize the genetic modification comprising a sequence from the pUB110 shuttle vector (GenBank: M19465.1), harbouring antimicrobial resistance genes conferring a resistance to kanamycine, neomycin and bleomycin, flanked on each side by a sequence coding for a protease (GenBank: WP_032874795.1). In addition, based on these data, two real-time PCR methods, that can be used by enforcement laboratories, specific to this unauthorized genetically modified bacterium were developed and validated. The present study emphasizes the key role that whole-genome sequencing can take for detection of unknown and unauthorized genetically modified microorganisms in commercialized microbial fermentation products intended for the food and feed chain. Moreover, current issues encountered by the Competent Authorities and enforcement laboratories with such unexpected contaminations and the importance of performing official controls were highlighted.

Integrated consolidated bioprocessing for simultaneous production of Omega-3 fatty acids and bioethanol

Lignocellulosic biomass represents a promising feedstock for supplying biofuels and other useful bioproducts in near future. As an example of process innovation, this study describes an integrated microbial fermentation process involving a thermoanaerobe and a marine thraustochytrid, aiming at the co-production of docosahexaenoic acids and bioethanol from rice straw biomass as substrate. Key points of the process are: (a) consolidated bioprocessing of pretreated rice straw biomass to bioethanol (anaerobic fermentation) resulting into 1.8 g L−1 bioethanol and 29.40% solubilization of rice straw biomass; (b) utilization of spent lignocellulose derived sugars in microalgal fermentation (aerobic fermentation) with subsequent promising cell growth (2.77 g L−1), substantial lipids (17.05%) and docosahexaenoic acids production (44.0% of total fatty acids). Other major fatty acids (as total fatty acid %) were palmitic acid (13.95%), stearic acid (5.07%), eicosapentaenoic acid (7.24%), and docosapentaenoic acid (16.12%). This study provides a new route for marine algae utilization of lignocellulosic biomass for co-production of bioethanol and value-added bioproducts.

Correction to: Production of poly hydroxy butyrate (PHB) from Eichhornia crassipes through microbial fermentation process

Correction to: Production of poly hydroxy butyrate (PHB) from Eichhornia crassipes through microbial fermentation process

Can Polyhydroxyalkanoates Be Produced Efficiently From Waste Plant and Animal Oils?

Polyhydroxyalkanoates (PHAs) are a potential replacement for some petrochemical-based plastics. PHAs are polyesters synthesized and stored by various bacteria and archaea in their cytoplasm as water-insoluble inclusions. PHAs are usually produced when the microbes are cultured with nutrient-limiting concentrations of nitrogen, phosphorus, sulfur, or oxygen and excess carbon sources. Such fermentation conditions have been optimized by industry to reduce the cost of PHAs produced commercially. Industrially, these biodegradable polyesters are derived from microbial fermentation processes utilizing various carbon sources. One of the major constraints in scaling-up PHA production is the cost of the carbon source metabolized by the microorganisms. Hence, cheap and renewable carbon substrates are currently being investigated around the globe. Plant and animal oils have been demonstrated to be excellent carbon sources for high yield production of PHAs. Waste streams from oil mills or the used oils, which are even cheaper, are also used. This approach not only reduces the production cost for PHAs, but also makes a significant contribution toward the reduction of environmental pollution caused by the used oil. Advancements in the genetic and metabolic engineering of bacterial strains have enabled a more efficient utilization of various carbon sources, in achieving high PHA yields with specified monomer compositions. This review discusses recent developments in the biosynthesis and classification of various forms of PHAs produced using crude and waste oils from the oil palm and fish industries. The biodegradability of the PHAs produced from these oils will also be discussed.

MEMO: A Method for Computing Metabolic Modules for Cell-Free Production Systems.

Cell-free bioproduction systems represent a promising alternative to classical microbial fermentation processes to synthesize value-added products from biological feedstocks. An essential step for establishing cell-free production systems is the identification of suitable metabolic modules with defined properties. Here we present MEMO, a novel computational approach to find smallest metabolic modules with specified stoichiometric and thermodynamic constraints supporting the design of cell-free systems in various regards. In particular, one key challenge for a sustained operation of cell-free systems is the regeneration of utilized cofactors (such as ATP and NAD(P)H). Given a production pathway with certain cofactor requirements, MEMO can be used to compute smallest regeneration modules that recover these cofactors with required stoichiometries. MEMO incorporates the stoichiometric and thermodynamic constraints in a single mixed-integer linear program, which can then be solved to find smallest suitable modules from a given reaction database. We illustrate the applicability of MEMO by calculating regeneration modules for the recently published synthetic CETCH cycle for in vitro carbon dioxide fixation. We demonstrate that MEMO is very flexible in taking into account the diverse constraints of the CETCH cycle (e.g., regeneration of 1 ATP, 4 NADPH and of 1 acetyl-group without net production of CO2 and with permitted side production of malate) and is able to determine multiple solutions in reasonable time in two large reaction databases (MetaCyc and BiGG). The most promising regeneration modules found utilize glycerol as substrate and require only 8 enzymatic steps. It is also shown that some of these modules are robust against spontaneous loss of cofactors (e.g., oxidation of NAD(P)H or hydrolysis of ATP). Furthermore, we demonstrate that MEMO can also find cell-free production systems with integrated product synthesis and cofactor regeneration. Overall, MEMO provides a powerful method for finding metabolic modules and for designing cell-free production systems as one particular application.

Fault diagnosis of microbial pharmaceutical fermentation process with non-Gaussian and nonlinear coexistence

A large Proportion of batch processes commonly have traits of non-Gaussian and nonlinear. In this work, Multiway Kernel Entropy Independent Component Analysis (MKEICA) algorithm was developed to formulate more accurate model for process monitoring so as to enhance the monitoring performance. The original process data with three-dimension were first expanded into two-dimensional data matrix by using AT variable expansion method. The Kernel Entropy Component Analysis (KECA) was then employed to preprocess the data in order to reduce data redundancy. Such approach can also retain the information of cluster structure and maximize the essential characteristics of data. After that, a monitoring model of MKEICA was established for production process monitoring. Once a fault is detected, a nonlinear contribution plots method would be utilized to diagnose the fault variables. Consequently, to illustrate the superiority and feasibility, the proposed method was conducted on the penicillin simulation platform and the actual pharmaceutical production process.

A strategy of untargeted foodomics profiling for dynamic changes during Fu brick tea fermentation using ultrahigh-performance liquid chromatography-high resolution mass spectrometry

During a microbial fermentation process, the chemical composition of Fu brick tea changed substantially. To better profile the unique changes of metabolites and lipids in Fu brick tea at different fermentation stages, a multifaceted strategy combining untargeted foodomics with UHPLC-Q-Orbitrap analysis was developed. The discriminative tea metabolites and lipids were determined by statistical analysis and online database matching. A total of 12 characteristic components (3 metabolic and 9 lipid variables) were found to differ significantly among samples. Quantitative analysis showed that, except for trimyristin and incensole acetate, the content of these compounds gradually increased and finally stabilized with increasing fermentation time. The simultaneous determination of metabolites and lipids in this study provided detailed information for the analysis of multi-component changes in a complex matrix.

Production of poly hydroxy butyrate (PHB) from Eichhornia crassipes through microbial fermentation process

Polyhydroxybutyrate (PHB) is one of the highly biodegradable and biologically acceptable thermoplastics synthesized by many microorganisms collectively called polyhydroxyalkanoates (PHAs). All available biopolymers are viewed as perfect answers for the resolution of natural contamination issue by supplanting ordinary plastic business. They are likewise utilized as osteosclerotic stimulants attributable to their piezoelectric properties, in bone plates, during operations as suture material and vein substitutions. Synthesis of PHB is found in a wide range of Gram’s negative and gram’s positive bacteria belonging to distinct genera. Optimum culture condition for the PHB producing microbes are provided, including restricted centralization of nitrogen, sulfur, phosphorus, or the trace elements and maximum convergence of carbon source Indeed, to market PHAs, significant exertion has been dedicated towards a decline in the production cost through the improvement of bacterial strains and enhancing effectiveness of recovery/fermentation procedure. This is being done considering the fact that substrate prices show the greatest impact on PHA's manufacturing cost. The price of the substrate used has the most significant influence on the production cost of PHA. In this research, a potential bacterial strain was isolated from the soil and tested for its PHB producing ability. The use of cheaper substrate for lowering the cost is prerequisite. For PHB production, water hyacinth was used as a carbon source. Bacterial growth was optimized for maximum PHB production. The optimum condition was found to be 30 °C, 8% substrate concentration and 72 h of incubation time.

Saccharomyces cerevisiae exhibiting a modified route for uptake and catabolism of glycerol forms significant amounts of ethanol from this carbon source considered as \u2018non-fermentable\u2019

BackgroundDue to its inevitable formation during biodiesel production and its relatively high degree of reduction, glycerol is an attractive carbon source for microbial fermentation processes. However, glycerol is catabolized in a fully respiratory manner by the eukaryotic platform organism Saccharomyces cerevisiae. We previously engineered S. cerevisiae strains to favor fermentative metabolism of glycerol by replacing the native FAD-dependent glycerol catabolic pathway with the NAD-dependent ‘DHA pathway’. In addition, a heterologous aquaglyceroporin (Fps1 homolog) was expressed to facilitate glycerol uptake. The current study was launched to scrutinize the formation of S. cerevisiae’s natural fermentation product ethanol from glycerol caused by the conducted genetic modifications. This understanding is supposed to facilitate future engineering of this yeast for fermenting glycerol into valuable products more reduced than ethanol.ResultsA strain solely exhibiting the glycerol catabolic pathway replacement produced ethanol at concentrations close to the detection limit. The expression of the heterologous aquaglyceroporin caused significant ethanol production (8.5 g L−1 from 51.5 g L−1 glycerol consumed) in a strain catabolizing glycerol via the DHA pathway but not in the wild-type background. A reduction of oxygen availability in the shake flask cultures further increased the ethanol titer up to 15.7 g L−1 (from 45 g L−1 glycerol consumed).ConclusionThe increased yield of cytosolic NADH caused by the glycerol catabolic pathway replacement seems to be a minimal requirement for the occurrence of alcoholic fermentation in S. cerevisiae growing in synthetic glycerol medium. The remarkable metabolic switch to ethanol formation in the DHA pathway strain with the heterologous aquaglyceroporin supports the assumption of a much stronger influx of glycerol accompanied by an increased rate of cytosolic NADH production via the DHA pathway. The fact that a reduction of oxygen supply increases ethanol production in DHA pathway strains is in line with the hypothesis that a major part of glycerol in normal shake flask cultures still enters the catabolism in a respiratory manner.

Optimization of Robust Model Predictive Controllers for Second-Order Biological Processes Using Markov Chain Monte Carlo Techniques

For biological fermentation processes in vaccine manufacturing, the development of a robust controller is essential to manage uncertainty in the process model and external disturbances in the control setup. A possible control strategy for this purpose is to use model predictive control (MPC), which utilizes a process model to compute a sequence of control moves via a receding horizon approach. To determine each control move, an MPC controller solves an optimization problem which can be modified to a min–max problem in order to achieve robustness. Extensive work has been undertaken to develop and test the min–max approach to obtain robust MPC; however, there is a paucity of control literature that has focused on the offline optimization of MPC control parameters (e.g., MPC horizons, control weights) to achieve robustness. To address this issue, a model predictive controller for a second-order process model representing a microbial fermentation process is developed. Uncertainty is simulated by generating 500 mismatched process models based on observed experimental data. The developed MPC controller is then applied to these process models, resulting in 500 scenarios of plant and nominal model mismatch, from which the average control performance and variation in control performance are computed. Markov Chain Monte Carlo (MCMC) techniques are successfully used to find the optimal settings that maximize control performance and minimize variation in control performance in response to step changes in both the set-point and the input disturbance. This novel approach may serve as a foundation for future MPC tuning studies which seek to determine the optimal combination of MPC settings that result in robust control.

Lactobacillus rhamnosus GG and Saccharomyces cerevisiae boulardii exert synergistic antipathogenic activity in vitro against enterotoxigenic Escherichia coli.

Short-term colonic in vitro batch incubations were performed to elucidate the possible synergistic effects of Lactobacillus rhamnosus GG (CNCM-I-4798) and Saccharomyces cerevisiae boulardii (CNCM-I-1079) (associated in Smebiocta/Smectaflora Protect®) on the colonic microbial fermentation process, as well as their antipathogenic activity against enterotoxigenic Escherichia coli (LMG2092) (ETEC). These incubations adequately simulate the native microbiota and environmental conditions of the proximal colon of both adult and toddler donors, including the colonic mucosal layer. Results indicated that both strains were capable of growing together without showing antagonistic effects. Co-cultivation of both strains resulted in increased butyrate (stimulated by L. rhamnosus GG), propionate (stimulated by S. boulardii), and ethanol (produced by S. boulardii) production compared to the control incubations, revealing the additive effect of both strains. After inoculation of ETEC under simulated dysbiotic conditions, a 40 and 46% reduction in the concentration of ETEC was observed upon addition of both strains during the experiments with the adult and toddler donor, respectively. Furthermore, ETEC toxin levels decreased upon S. boulardii inoculation, probably due to proteolytic activity of this strain, with a synergistic effect being observed upon co-cultivation of L. rhamnosus GG and S. boulardii resulting in a reduction of 57 and 46% for the adult and toddler donor, respectively. Altogether, the results suggest that both probiotics together may help microbiota functionality, in both adults and toddlers and under healthy or impaired conditions, which could be of great interest when the colonic microbiota is dysbiotic and therefore sensitive to pathogenic invasion such as during antibiotic treatment.

Robust parameter estimation for constrained time-delay systems with inexact measurements

<p style='text-indent:20px;'>In this paper, we consider estimation problems involving constrained nonlinear systems with the unknown time-delays and unknown system parameters. These unknown quantities are to be estimated such that a least-squares error function between the system output and a set of noisy measurements is minimized subject to the characteristic time constraints specifying the restrictions. We first present the classical estimation formulation, where the expectation of the error function is regarded as the cost function. Then, in order to obtain robust estimates against the noises in measurements, we propose a robust estimation formulation, in which the cost function is the variance of the error function and an additional constraint indicates an allowable sacrifice from the optimal expectation value of the classical estimation problem. For these two estimation problems, we derive the gradients of the corresponding cost and constraint functions with respect to time-delays and system parameters by solving some auxiliary time-delay systems backward in time. On this basis, we develop gradient-based optimization algorithms to determine the optimal time-delays and system parameters. Finally, we consider two example problems, including a parameter estimation problem in microbial batch fermentation process, to illustrate the effectiveness and applicability of our proposed algorithms.

Study on the Solubility between Diesel and Acetone–Butanol–Ethanol with or without Water

Acetone–butanol–ethanol (ABE) is an intermediate product in the microbial fermentation process for producing biobutanol. ABE not only retains the advantages of oxygen-containing fuels but also redu...

High precision qualitative identification of yeast growth phases using molecular fusion spectra

Compared to a single molecule spectral analysis method, molecular fusion spectroscopy can obtain multi-dimensional information reflecting the target attributes more comprehensively, which makes this method have potential application value in online monitoring of microbial fermentation process. This study proposes a method based on near-infrared (NIR) and Raman molecular fusion spectroscopy for high precision identification of yeast growth phases. First, the original spectra of yeast broth were preprocessed using Savitzky-Golay (SG) convolution smoothing combined with standard normal variate (SNV). Then, a novel fusion algorithm, which was discriminant correlation analysis (DCA), was used to realize the effective fusion of the NIR and Raman spectra after preprocessing in the feature layer. Finally, a linear discriminant analysis (LDA) identification model was developed based on the DCA fusion feature vectors. The results showed that the prediction performance of the LDA identification model based on the eigenvectors by the DCA fusion is far better than the best LDA model based on the single molecule spectra, and its correct identification rate is 97.37% in the prediction set. The overall results demonstrate that molecular fusion spectroscopy can achieve high precision and rapid monitoring of yeast fermentation process, and the DCA is an effective tool for molecular spectral fusion.

Microbial industrial enzymes: An annotated selection of World Wide Web sites relevant to the topics in microbial biotechnology.

https://link.springer.com/chapter/10.1007/978-3-319-52287-6_28 This review article in an e-book provides an excellent overview to the field of industrial enzymes. In addition to the current commonly used enzymes, it contains information of emerging enzymes of interest. http://www.novozymes.com/-/media/Project/Novozymes/.../Enzymes_at_work.pdf This web brochure produced by Novozymes provides many examples of industrial enzymes, their origins and applications. http://www1.lsbu.ac.uk/water/enztech/detergent.html This web article is a little dated but it nevertheless provides a good overview of how enzymes are used on washing formulations. https://www.aasv.org/shap/issues/v18n2/v18n2p90.html Phytase is an enzyme that releases phosphate from phytate, a hexaphosphate ester of inositol. Because much phosphate in grain and seeds is stored in phytate, phytase enzyme is added to animal feed to help release phosphate from the feed. https://www.ocl-journal.org/articles/ocl/pdf/2017/04/ocl170015.pdf Lipases, mostly obtained from fungi, is used extensively in industry. Major uses are in detergents and in food oil processing. https://www.heraproject.com/files/22-F-07_PROTEASE_HERA_Final%20Edition%20(unsecured%20-%20PDFA-1b).pdf Subtilisin is a protease, originally obtained from strains of Bacillus. It is used in laundry, dishwashing and contact lens cleaning applications. The current document contains much information relevant to the industrial use of subtilisin. https://www.chemistryworld.com/podcasts/chymosin/3007682.article Chymosin is an important enzyme used in the conversion of milk to cheese. It was originally obtained from the digestive tracts of young ruminant animals. More recently, it is produced in recombinant form by microbial fermentation processes. https://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-016-0507-6 Cellulases are important industrial enzymes in biomass conversion. A major organism long studied for it enzymes for degrading cellulose is the fungus Trichoderma reesei. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3769773/ This review article discusses the uses of amylases, mostly in converting various food products such as starches in corn, wheat, rice, and tapioca. A very large process currently is the conversion of corn starch to glucose by amylase coupled to the isomerization of glucose to fructose. https://www.researchgate.net/publication/315803185_Xylanases_and_their_industrial_applications_A_review This Research Gate website contains an abstract on xylanases and a link to the full review article on the topic. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3347563/ This review article covers enzymes used in the production of semi-synthetic beta-lactam antibiotics. The major focus is on protein expression and engineering. https://www.creative-enzymes.com/similar/glucose-isomerase_319.html Glucose isomerase, also known as xylose isomerase, is widely used in the production of high-fructose corn syrup. https://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-biology/pectinase Pectinases are widely used in producing clarified fruit juices. The pectinases hydrolyze linkages in the pectic substance found in the cell walls of plants. https://www.novozymes.com/en Novozymes is a major enzyme producing company, headquartered in Denmark. http://biosciences.dupont.com DuPont is a very large producer of enzymes with currently 900 commercial products on the market. https://www.dupont.com/products-and-services/industrial-biotechnology/industrial-enzymes-bioactives/primagreen.html Many enzymes are used in textile processing and clothing finishing. This website addresses are group of enzymes used in those applications. http://animalnutrition.dupont.com/productsservices/feed-enzymes/ Animal nutrition enzymes are largely derived from bacteria and aid in the animal digestion of their foods. The enzymes are principally phytase, proteases, and carbohydrate hydrolyzing enzymes. https://www.basf.com/global/en/products/cross-industry-solutions/enzymes.html The German chemical company BASF made a large entry into industrial enzymes with the the purchase of the Verenium Corporation. https://www.amano-enzyme.com/uk/about-company/ Amano produces microbial enzymes for use in food, supplement, and medical diagnostic industries.