Bioprocess of Gibberellic Acid by Fusarium fujikuroi: The Challenge of Regulation, Raw Materials, and Product Yields.

Gibberellic acid (GA3) is a tetracyclic diterpenoid carboxylic acid synthesized by the secondary metabolism of Fusarium fujikuroi. This phytohormone is widely studied due to the advantages it offers as a plant growth regulator, such as growth stimulation, senescence delay, flowering induction, increased fruit size, and defense against abiotic or biotic stress, which improve the quality and yield of crops. Therefore, GA3 has been considered as an innovative strategy to improve agricultural production. However, the yields obtained at large scale are insufficient for the current market demand. This low productivity is attributed to the lack of adequate parameters to optimize the fermentation process, as well as the complexity of its regulation. Therefore, this article describes the latest advances for potentializing the GA3 production process, including an analysis of its origins from crops, the benefits of its application, the related biosynthetic metabolism, the maximum yields achieved from production processes, and their association with genetic engineering techniques for GA3 producers. This work provides a new perspective on the critical points of the production process, in order to overcome the limits surrounding this modern line of bioengineering.

Phytochemical profile of petals from black Dahlia pinnata by flow injection analysis-electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry.

INTRODUCTION: Dahlia pinnata Cav. is a flower native to Mexico that has many applications; in particular, its petals have been used for ornamental, food, and medicinal purposes, for example to treat skin rashes and skin cracks. It has been reported that the medicinal properties of plants are generally related to the phytochemical constituents they possess. However, there are few studies on black D. pinnata. OBJECTIVES: The present study was aimed at qualitatively and quantitatively determining the phytochemical profile of petals from black D. pinnata. METHODOLOGY: Phytochemicals from Dahlia petals were extracted by consecutive maceration (hexane, dichloromethane, and methanol); then, the extracts were analyzed through colorimetric assays and UV-Vis spectroscopy for qualitative identification and quantification of phytochemical compounds, respectively. The methanolic extract was analyzed by flow injection analysis-electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry (FIA-ESI-FTICR-MS) in negative and positive mode. RESULTS: Quantitative phytochemical profiling of the methanolic extract by UV-Vis spectroscopy indicated high contents of phenolic compounds (34.35 ± 3.59 mg EQ/g plant) and sugars (23.91 ± 1.99 mg EQ/g plant), while the qualitative profiling by FIA-ESI-FTICR-MS allowed the tentative identification of several flavonoids and phenolic acids. Kaempferol-3-rutinoside, pelargonidin-3-(6″-malonylglucoside)-5-glucoside, rutin, kaempferol-3-(2″,3″-diacetyl-4″-p-coumaroylrhamnoside), and myricetin-3-(2‴-galloylrhamnoside) were the main compounds detected. CONCLUSION: The results expand our knowledge of the phytochemical constituents of petals from black D. pinnata.

Cannabis Extraction Technologies: Impact of Research and Value Addition in Latin America.

The Cannabis genus of plants has been widely used in different cultures for various purposes. It is separated into three main species: sativa, indica, and ruderalis. In ancient practices, the plant was used as a multipurpose crop and valued for its fiber, food, and medicinal uses. Since methodologies for the extraction, processing, and identification of components have become available, medical, and food applications have been increasing, allowing potential development in the pharmaceutical and healthy functional food industries. Although the growing legalization and adoption of cannabis for the treatment of diseases are key factors pushing the growth of its market, the biggest challenge is to obtain higher-quality products in a time- and cost-effective fashion, making the process of extraction and separation an essential step. Latin American countries exhibit great knowledge of extraction technologies; nevertheless, it is still necessary to verify whether production costs are economically profitable. In addition, there has been an increase in commercial cannabis products that may or may not be allowed, with or without quality fact sheets, which can pose health risks. Hence, legalization is mandatory and urgent for the rest of Latin American countries. In this article, the phytochemical compounds (cannabinoids, terpenes, and phenolic compounds), the current status of legalization, extraction techniques, and research advances in cannabis in Latin America are reviewed.

Two-methods approach to follow up biomass by impedance spectroscopy: Bacillus thuringiensis fermentations as a study model.

Impedance spectroscopy is used for the characterization of electrochemical systems as well as for the monitoring of bioprocesses. However, the data obtained using this technique allow multiple interpretations, depending on the methodology implemented. Hence, it is necessary to establish a robust methodology to reliably follow-up biomass in fermentations. In the present work, two methodological approaches, mainly used for the characterization of electrochemical systems, were employed to characterize and determine a frequency that allows the monitoring of biomass in Bacillus thuringiensis fermentations by impedance spectroscopy. The first approach, based on a conventional analysis, revealed a single distribution with a characteristic frequency of around 2 kHz. In contrast, the second approach, based on the distribution of relaxation times, gave three distributions (A, B, and C). The C distribution, found near 9 kHz, was more related to the microbial biomass than the distribution at 2 kHz using the equivalent circuits. The time course of the B. thuringiensis fermentation was followed; bacilli, spores, glucose, and acid and base consumption for pH were determined out of line; and capacitance at 9 kHz was monitored. The correlation between the time course data and the capacitance profile indicated that the monitoring of B. thuringiensis at 9 kHz mainly corresponds to extracellular activity and, in a second instance, to the cellular concentration. These results show that it is necessary to establish a robust and reliable methodology to monitor fermentation processes by impedance spectroscopy, and the distribution of relaxation times was more appropriate. KEY POINTS: • Application of impedance spectroscopy for bioprocess monitoring • Low-frequency monitoring of biomass in fermentations • Analysis of impedance data by two methodological approaches.

On-line monitoring of industrial interest Bacillus fermentations, using impedance spectroscopy.

Impedance spectroscopy is a technique used to characterize electrochemical systems, increasing its applicability as well to monitor cell cultures. During their growth, Bacillus species have different phases which involve the production and consumption of different metabolites, culminating in the cell differentiation process that allows the generation of bacterial spores. In order to use impedance spectroscopy as a tool to monitor industrial interest Bacillus cultures, we conducted batch fermentations of Bacillus species such as B. subtilis, B. amyloliquefaciens, and B. licheniformis coupled with this technique. Each fermentation was characterized by the scanning of 50 frequencies between 0.5 and 5 MHz every 30 min. Pearson's correlation between impedance and phase angle profiles (obtained from each frequency scanned) with the kinetic profiles of each strain allowed the selection of fixed frequencies of 0.5, 1.143, and 1.878 MHz to follow-up of the fermentations of B. subtilis, B. amyloliquefaciens and B. licheniformis, respectively. Dielectric profiles of impedance, phase angle, reactance, and resistance obtained at the fixed frequency showed consistent changes with exponential, transition, and spore release phases.

Use of equivalent circuit analysis and Cole-Cole model in evaluation of bioreactor operating conditions for biomass monitoring by impedance spectroscopy.

The most important parameter in bioprocesses is biomass, where not only the quantity produced in a culture but also the behavior that is presented are important concerns. It is clear that conditions of operation in a bioreactor affect biomass production, but how operation conditions affect the measurement of biomass on-line is of special interest. We studied the effect of bioreactor operating condition variations on model parameters using impedance spectroscopy for biomass monitoring. The model parameters analyzed were capacitance, resistance, alpha (α), conductivity delta (∆σ) and critical frequency (fc). These model parameters were obtained by fitting data from impedance measurements to an equivalent circuit model and Cole-Cole conductivity model. The effect of operating conditions on the medium with no cells was estimated by the percentage of change in each model parameter. The operating conditions with the most significant percentage of change were determined, by comparing to the percentage of change of the same model parameters obtained, during a fermentation of Bacillus thuringiensis as a cellular model. Equivalent circuit parameters were mainly affected by variation in pH, temperature and aeration, whereas Cole-Cole parameters were affected by variation in agitation, aeration, temperature and pH. Therefore, any variation in these operating conditions (within the test interval) during a fermentation may generate changes in monitoring parameters, which will not be a direct consequence of any change in the properties of the biomass.