TL;DR
Overall, the use of metagenomics, artificial intelligence, and experimental approaches has tremendous potential for developing efficient and economically viable lipases for sustainable biotechnological applications.
Microbial inulinases are increasingly recognized as valuable biocatalysts for the sustainable production of high-value products, including fructooligosaccharides, fructose, bioethanol, and organic acids in industries, such as food, pharmaceuticals, and bioenergy. In the last few decades, microbial inulinase research has advanced significantly, from strain selection and fermentation optimization to advanced enzyme engineering and immobilization, improving yields, stability, and reusability. There are still some final bottlenecks, such as low yields, poor thermostability, and high purification costs. This review examines strategies to innovate and overcome these bottlenecks, including novel immobilization strategies that utilize nanomaterials, system-scale bioprocess optimization using artificial intelligence (AI), and bioprospecting extremophiles using metagenomics. The present review discusses how statistical and computational modeling (RSM, ANN, and AI) significantly increases yield and process efficiency, with comments on their relevance to contemporary biorefinery applications. The advanced immobilization approaches significantly enhance operational stability and reusability, allowing for continuous processing. This review situates the development of inulinase as not just an enzymological effort but a multidisciplinary effort involving process engineering and sustainability science. Overall, emphasize is given toward the thought that advancements leaning toward the future will require a synthesis of AI-designed enzyme systems; economical immobilization supports; and incorporation of circular bioeconomy principles through the valorization of agro-wastes. These barriers to knowledge transfer must be resolved if we are to unlock the full bioeconomic potential of microbial inulinase systems.
Oil pollution remains a persistent global environmental challenge due to the recalcitrance and toxicity of lipid-rich contaminants in terrestrial and aquatic ecosystems. Bacterial lipases (EC 3.1.1.3) play a pivotal role in the initial stages of bioremediation by catalysing the hydrolysis of complex lipids into more bioavailable intermediates, thereby facilitating downstream microbial degradation and mineralisation. This review critically examines the mechanistic basis of lipase-mediated hydrocarbon degradation, with emphasis on enzyme structure–function relationships, catalytic pathways, and regulation under environmentally relevant conditions. In addition to conventional applications in soil and wastewater bioremediation, emerging strategies involving immobilised enzymes, microbial consortia, and waste-derived substrates are evaluated for their effectiveness and scalability. Attention is given to advances in molecular and omics approaches, including metagenomics, transcriptomics, and proteomics, which have expanded the discovery of novel lipases but remain limited in their ability to predict in situ functionality. The review highlights the growing role of protein engineering and artificial intelligence in tailoring lipase properties; however, it also critically assesses current limitations, including insufficient experimental validation and challenges in translating computational predictions to complex environmental systems. Furthermore, integrating multi-omics data into quantitative and predictive frameworks is identified as a key future direction for improving bioremediation efficiency. Despite significant progress, major gaps persist in linking enzyme activity to real-world degradation performance and in developing standardized, scalable approaches. This review therefore provides a comprehensive and critical synthesis of current knowledge while identifying strategic research priorities required to advance bacterial lipases as robust tools for sustainable bioremediation of lipid-based pollutants.
Microbial lipases are indispensable in modern environmental management and industry, offering eco-friendly, efficient, and costeffective solutions across diverse sectors. Their broad specificity, high stability, and ability to catalyse reactions under mild conditions underscore their significance in promoting sustainable development and innovative industrial processes. As biotechnological advancements continue to garner momentum, the potential applications of lipases are expected to expand, further enhancing their role in shaping a more sustainable and environmentally conscious future. Lipases are universal enzymes of extensive physiological importance. They have a wide range of applications across industries, including food (dairy, fats and oils, beverages, and bakery), detergents, paper and pulp, biodiesel production, pesticides, pharmaceuticals, and bioremediation of various effluents. Microorganisms producing lipase are derived primarily from bacterial genera such as Bacillus, Pseudomonas, and Burkholderia, as well as fungal species such as Rhizopus, Aspergillus, and yeast-like Candida, enabling efficient production from diverse substrates, including agro-industrial wastes. Their broad substrate specificity, stability under extreme conditions, and ability to catalyze reactions such as hydrolysis, esterification, and transesterification underpin transformative applications across sectors. As global industries increasingly shift toward green technologies and sustainable processes, microbial lipases are expected to play an even more crucial role in addressing environmental challenges while supporting innovation and industrial development.
Candida yeasts represent a versatile yet underexploited platform for industrial biotechnology. These yeasts utilize a remarkably broad range of carbon sources, particularly for hydrophobic carbon sources, coupled with robust growth and diverse biosynthetic capacities, making them promising hosts for sustainable production of chemicals, fuels, and proteins. Despite these advantages, industrial deployment of Candida species has been hindered by concerns regarding opportunistic pathogenicity and the historical lack of efficient genetic manipulation tools, leading to a substantial gap between metabolic potential and practical utilization. Recent advances in functional genomics, genome editing, and systems metabolic engineering are rapidly overcoming these barriers, enabling more precise and efficient strain development. In this review, we systematically summarize recent progress in the metabolic engineering of Candida species as microbial cell factories, with particular emphasis on expanding genetic toolkits, utilizting renewable and non-conventional carbon sources, and biosynthesizing high-value compounds. In addition, we propose a biosafety-oriented classification framework to support their safe industrial deployment. Finally, we discuss current challenges and emerging opportunities, emphasizing that the synergy of synthetic biology and artificial intelligence-driven design holds the key to unlocking the biotechnological potential of Candida yeasts.
Lipases, versatile enzymes produced by microbes, find extensive applications in diverse industrial processes, with notable significance in the laundry industry where lipase-based detergents efficiently eliminate lipid-based stains from textiles, enhancing cleanliness and sustainability. The utilization of microbial lipases in detergent formulations offers significant potential for reducing environmental impact. Marine environments, renowned for their extreme conditions, harbor microorganisms with robust enzymatic capabilities, making them attractive candidates for enzyme production. In this study, lipase enzymes sourced from marine bacteria, particularly Brevibacillus borestelensis, were targeted for production and partial purification to assess their suitability as detergent additives. Through process optimization, lipase production was enhanced, and subsequent partial purification facilitated the evaluation of enzyme stability and activity in the presence of various commercially available detergents. The identification of the marine bacteria was achieved through sequencing the 16S rRNA gene, confirming their taxonomic classification. The purified lipase exhibited a molecular mass of approximately 35 kDa and a specific activity of 110.8 U/mg. Furthermore, the stability of the lipase with commonly used detergent powders was evaluated, demonstrating compatibility with various detergents, with more than 80% relative activity retained. These findings underscore the potential of microbial lipases, particularly those sourced from extreme environments, as environmentally friendly additives in detergent formulations, contributing to sustainable cleaning practices in the industry.
Artificial enzymes represent transformative biocatalysts that overcome the inherent limitations of natural enzymes in substrate scope and activity specificity, enabling challenging new-to-nature reactions. Despite remarkable advances in designing diverse artificial enzymes through synthetic cofactor or non-canonical amino acid incorporation, their widespread industrial application remains hindered by the high costs and operational complexity of in vitro systems. Whole-cell biocatalysis emerges as a promising alternative by capitalizing on the cellular environment, including cofactor regeneration, multi-enzyme cascades, and enhanced enzyme stability. This review provides a comprehensive introduction to recent progress in artificial enzyme design and their successful applications in whole-cell biocatalysis. We summarize innovative protein engineering tools for creating artificial enzymes, assembly strategies to enhance in vivo catalytic performance, and representative applications of artificial enzymes-containing whole-cell systems in non-natural transformations. We also discuss the challenges and prospects of whole-cell artificial enzyme catalysis in advancing sustainable and scalable biomanufacturing.
