Review
Temperature-resistant and solvent-tolerant lipases as industrial biocatalysts: Biotechnological approaches and applications

https://doi.org/10.1016/j.ijbiomac.2021.07.101Get rights and content

Highlights

  • TR and ST lipases gain a remarkable interest for industrial applications.

  • There are some features distinguishing TR and ST lipases.

  • DNA recombination techniques are commonly used to enhance the lipase productivity.

  • Protein engineering approaches are ordinarily applied to increase the lipase stability.

  • Immobilization of lipase also used to improve the lipase stability.

Abstract

The development of new biocatalytic systems to replace the chemical catalysts, with suitable characteristics in terms of efficiency, stability under high temperature reactions and in the presence of organic solvents, reusability, and eco-friendliness is considered a very important step to move towards the green processes. From this basis, the use of lipase as a catalyst is highly desired for many industrial applications because it offers the reactions in which could be used, stability in harsh conditions, reusability and a greener process. Therefore, the introduction of temperature-resistant and solvent-tolerant lipases have become essential and ideal for industrial applications. Temperature-resistant and solvent-tolerant lipases have been involved in many large-scale applications including biodiesel, detergent, food, pharmaceutical, organic synthesis, biosensing, pulp and paper, textile, animal feed, cosmetics, and leather industry. So, the present review provides a comprehensive overview of the industrial use of lipase. Moreover, special interest in biotechnological and biochemical techniques for enhancing temperature-resistance and solvent-tolerance of lipases to be suitable for the industrial uses.

Introduction

The discovery of practical solutions for clean, green and ecofriendly large-scale industrial processes of has become a pressing necessity [1]. There is a great demand for the transition from chemical to biological catalysts that empower green processes, involving utilization of bio-products. The continuously growing interest in lipases stems from their biotechnological adaptability and capability to catalyze a wide variety of bioconversion reactions in different fields, such as food, pharmaceutical, biofuel, biomedical, and chemical industries [2].

Most of the currently available conventional enzymes are mesophilic in nature, originating from microbes that grow optimally at moderate temperatures, i.e., ranging from 20 to 45 °C. Nonetheless, these microbes exhibit optimal behavior within a limited range of conditions and exhibit poor stability at elevated temperatures or in organic solvents. Generally, they underperform in industrial settings, which are often harsh for biosystems, e.g., use of protein denaturing solvents and high pressure in most industrial processes [3]. Hence, there is a great need for more efficient and resistant biocatalysts, which may lead to advances in the field of engineering mesophilic enzymes. Therefore, the development of temperature-resistant (TR) and solvent-tolerant (ST) enzymes is a key step towards improved biocatalysts [4].

Lipases produced from extremophiles revealed reasonable applicability in commercial bioprocesses and biotransformations due to their exceptional stability in organic solvents and high temperatures [5]. Plants, and animal are important sources of industrial lipases however, microorganisms are considered the most powerful, diverse and suitable source of TR and ST lipases. Microbial extremophilic lipases especially TR and ST enzymes, are strongly recommended due to their easy production, separation, and ability to undergo molecular modification [6]. Furthermore, TR lipases exhibit high reaction rates during purification and refining steps and are easily separated from other heat-unstable biomolecules [7], [8], [9].

TR and ST lipases are not dissimilar to their mesophilic relatives, in respect to structural conformation. Each protein has its own molecular mechanism of high temperature-resistance like the presence of disulfide bonds, hydrogen bonds, salt bridges, hydrophobic interactions, aromatic interactions, and metal binding [10], [11]. With breakthroughs in biotechnology many lipases have been designed to meet the industrial requirements [12]. Various molecular techniques (genomic recombination and protein engineering) were also performed to enhance the performance and efficiency of microbial lipases for their broader use in a variety of industries. As a result, many value-added products are being synthesized in the global market with the use of established technology employing purposely engineered lipases [13].

Many of these applications utilize immobilized lipases. The immobilization method has several advantages, such as improved biocatalyst thermal stability, withstanding in organic solvent media, and easy separation of lipases from the reaction media after repeated use [14], [15], [16].

As mentioned previously, the immobilization of lipases is important for their use in industrial-scale reactions. Therefore, the support materials employed for lipase immobilization should possess features required for proper biocatalytic systems, such as stability under high temperature, high surface area, high affinity to lipases, low cost, and easy availability. Moreover, the selection of the immobilization procedure robustly provide the desired properties necessary for the biocatalytic system according to the reaction type [17], [18], [19], [20].

This review highlights the using of TR and ST lipases in different industrial uses. In addition, the general features that distinguish lipolytic microbes capable of producing TR and ST lipases are also summarized. Besides, the structure-function relationship of lipases and their mechanism of action that catalyzes biodiesel production for example are discussed. Many lipases need structural modifications to become suitable for industrial usage. Therefore, biotechnological approaches applied to improve lipase performance; genomic recombination and protein engineering, as well as the recent advances in the immobilization of lipases were surveyed. Moreover, the many advantages and illustrations of the utilization of TR and ST lipases for applying in different applications and biodiesel production have been extensively reviewed.

Section snippets

Structural aspects and catalytic mechanism of lipases

Despite their low sequence identity, lipases exhibit a similar fold. Lipases belong to the α-β hydrolase family and have common structural characteristics with proteases, peroxidases, esterases, dehalogenases, and epoxide hydrolases [21]. The three-dimensional structural fold of α hydrolases has a central β-sheet containing eight analogous β-strands with the exclusion of β2 strand, which is oriented in the opposite direction to the other strands. Moreover, the central β-sheet comprises of a

TR lipases

Lipases are pervasive in nature and catalyze the breakdown of the ester linkage of acylglycerols containing long-chain fatty acids in aqueous media. They exhibit fascinating and marvelous features that make them outstanding biocatalysts, including stereo-, regio-, and enantio-selectivity, differential substrate specificities, and the probability of catalyzing mixed-phase reactions in aqueous media [40], [41]. The constant alterations in the three-dimensional structure and the loss of activity

Role of organic solvents in enzymatic transesterification for biodiesel production

The dissolution of reactants of the same polarity improves with the addition of organic solvents. Additionally, organic solvents minimize the reaction viscosity, which improves substrate diffusion to the active sites of the enzyme [83], [84]. This increases the rate of reaction and decreases enzyme inhibition. Several moderately priced organic solvents can also be used for this purpose. However, the main characteristic that is considered during solvent selection is their hydrophobicity [85],

Approaches for enhancing temperature resistance and solvent tolerance of lipases

Different strategies are used to enhance the stability of lipases in high-temperature reactions, to improve their activity in the presence of organic solvents for biodiesel production, and for other industrial purposes [109]. The most common and successful methods used for this purpose are: (i) increasing the efficiency of lipases by applying gene engineering techniques, especially genomic recombination (expression of the wild gene in a host strain) [110]; (ii) modifying amino acids in the

Biodiesel production

Biodiesel derived from triglycerides is one of the best alternate energy sources and has gained considerable scientific attention. It holds the promise to compensate for the increasing demand of petro-diesel [180], [181], [182], [183].

Biodiesel can be produced by several methods; however, the most practical and efficient method of biodiesel production thus far is the transesterification of triglycerides to produce long alkyl-chain esters and glycerol. This reaction requires chemical or

Conclusion and future perspectives

The use of TR and ST lipases as biocatalysts for industrial application has become a topic of great interest. Because of their natural and/or modified structural features, they can facilitate diverse types of chemical reactions at high temperatures and in presence of organic solvents for achieving high product yield, especially on an industrial scale. DNA recombination and protein engineering methods are essential tools to modify lipase structure at the molecular scale to provide lipases the

Funding

This work was supported by the National Research Foundation of Korea, Republic of Korea [grant number NRF-2019R1F1A1052625].

Declaration of competing interest

Authors declare that there is no conflict of interest.

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