Your browser does not fully support modern features. Please upgrade for a smoother experience.
Submitted Successfully!
Thank you for your contribution! You can also upload a video entry or images related to this topic. For video creation, please contact our Academic Video Service.
Version Summary Created by Modification Content Size Created at Operation
1 ADAMU MATINJA IDRIS -- 2276 2023-01-05 08:23:44 |
2 layout Sirius Huang -3 word(s) 2273 2023-01-06 10:37:42 |

Video Upload Options

We provide professional Academic Video Service to translate complex research into visually appealing presentations. Would you like to try it?
No, upload directly Yes
Cite
If you have any further questions, please contact Encyclopedia Editorial Office.
Matinja, A.I.;  Kamarudin, N.H.A.;  Leow, A.T.C.;  Oslan, S.N.;  Ali, M.S.M. Cold-Active Lipases and Esterases. Encyclopedia. Available online: https://encyclopedia.pub/entry/39776 (accessed on 08 February 2026).
Matinja AI,  Kamarudin NHA,  Leow ATC,  Oslan SN,  Ali MSM. Cold-Active Lipases and Esterases. Encyclopedia. Available at: https://encyclopedia.pub/entry/39776. Accessed February 08, 2026.
Matinja, Adamu Idris, Nor Hafizah Ahmad Kamarudin, Adam Thean Chor Leow, Siti Nurbaya Oslan, Mohd Shukuri Mohamad Ali. "Cold-Active Lipases and Esterases" Encyclopedia, https://encyclopedia.pub/entry/39776 (accessed February 08, 2026).
Matinja, A.I.,  Kamarudin, N.H.A.,  Leow, A.T.C.,  Oslan, S.N., & Ali, M.S.M. (2023, January 05). Cold-Active Lipases and Esterases. In Encyclopedia. https://encyclopedia.pub/entry/39776
Matinja, Adamu Idris, et al. "Cold-Active Lipases and Esterases." Encyclopedia. Web. 05 January, 2023.
Cold-Active Lipases and Esterases
Edit
This entry is adapted from the peer-reviewed paper 10.3390/ijms232315394

One of the survival strategies adopted by microorganisms living in cold environments is their expression of cold-active enzymes that enable them to perform an efficient metabolic flux at low temperatures necessary to thrive and reproduce under those constraints. Cold-active enzymes are ideal biocatalysts that can reduce the need for heating procedures and improve industrial processes’ quality, sustainability, and cost-effectiveness.

cold adaptation esterase lipase psychrophilic enzymes purification

1. Introduction

Cold-active enzymes are produced by psychrophilic microorganisms that are often heat-labile and perform a high catalytic activity at moderate to very low temperatures in contrast to their thermophilic and mesophilic orthologs [1][2]. The cold-active extremozymes generally achieved their efficient biochemical reactions by lowering both the enthalpy of activation and Gibbs free energy compared to their thermophilic and mesophilic counterparts [3]. Cold-active enzyme structures are homologous to their mesophilic counterparts. They only differ by discrete changes in their amino acid and spatial polypeptide structures, which are responsible for their distinct functions [4][5].
Cold-active enzymes have a high specific activity, low-affinity on the substrate at low temperatures, and they are structurally more flexible at their active sites; this flexible nature is due to weak intermolecular forces and increased exposure of hydrophobic residues [3][6][7]. Compared to their mesophilic and thermophilic counterparts, these features of high catalytic efficiency at low temperatures make these extremozymes highly attractive to the scientific community and provide potential applications in detergency, bioremediation, biofuels, and food industries [2][8][9]. Cold-active hydrolases such as protease, lipase, amylase, and cellulase were the most frequent enzymes characterised and used for industrial purposes compared to other cold-active enzymes [10][11][12][13]. Cold-active enzymes, in general, are ideal biocatalysts that can reduce the need for heating procedures, which improves the sustainability, cost-effectiveness, energy consumption, and quality of industrial production [2].

2. Cold-Active Lipase and Esterase Overexpression in Recombinant Heterologous Hosts

The most common strategy for obtaining large quantities of desired proteins is recombinant overexpression in a heterologous host [14][15]. Although the technique is often used in producing cold-active lipases and esterases, it is not specific to even cold-adapted enzymes but all recombinant proteins. When expressed in the cytosol, recombinant proteins are often produced at a greater yield, but they may also be regulated to be released into the culture media [14]. The overexpression of recombinant cold-active lipase and esterase is often achieved using mesophilic expression systems such as E. coli [16], yeast [17], and insects [18]. The production of large quantities of such enzymes at high concentrations remains challenging. As for other cold-active enzymes, the temperatures that cold-active lipase and esterase require for proper folding is inconsistent with the optimal growth temperature of these expression hosts [2]. The typical approach to mitigate folding problems in E. coli is to reduce the post-induction temperature below 20 °C. However, this slows down the host growth rate and the heterologous enzyme’s synthesis rate. Table 1 summarises some recently reported overexpression of cold-active lipase and esterase in a recombinant heterologous host.
E. coli was selected as the preferred expression host, and just one of the enzymes was produced in Saccharomyces cerevisiae (S. cerevisiae). However, the E. coli Rosetta TM strain was reported to be used once [19], BL21 (DE3), was the most popular. Other Gram-negative bacteria, such as Pseudomonas and Burkholderia, lack suitable promoters and require foldase (a special chaperon) and extracellular fatty acids to induce their expression, a mechanism that is primarily unclear [20][21]. The two most common yeasts used for expression systems were S. cerevisiae (Baker’s yeast) and Pichia pastoris (P. pastoris). Its major drawback is its strong natural tendency of S. cerevisiae to ferment carbohydrates to ethanol, which is toxic at low culture density. However, P. pastoris lacks the problem of harmful ethanol synthesis, but it cannot express any gene of interest. While specific proteins may have no issues with being expressed, others may have problems associated with glycosylation, secretion, and folding [22][23]. A recent study on recombinant overexpression by Xue, Yao [17] found excellent expression of cold-active esterase in the S. cerevisiae heterologous host, which was attributed to similarities between the yeast family to which the wild gene and S. cerevisiae belongs. Since the carbon source was n-propanol and isobutanol and not sugars, the limitation of using S. cerevisiae was not mentioned. Another heterologous host for recombinant proteins is insect cell culture systems, which are well-known for their use in creating vaccines and viral insecticides [24][25]. Compared to other eukaryotic expression systems, high levels of heterologous gene expression are frequently achieved, especially for intracellular proteins [26]. In several instances, the recombinant proteins are soluble and easily collected from infected cells [26][27]. In one study, a Yarrowia lipolytica (LIPY8) extracellular lipase gene was expressed using a baculovirus expression system in insect cells, and it was interesting that the best pH and temperature for cold-active lipase LipY8p expressed in insect cells were very different from those for the same enzyme expressed in P. pastoris [18]. Moreover, it is too early to conclude how the change in heterologous host from yeast to insect increases the cold activeness of a particular enzyme. On the other hand, adaptability to a wide range of culture broths and its rapid growth and high enzyme yield were the major favourable characteristics that allowed the utilisation of E. coli for recombinant overexpression of heterologous proteins [28][29]. The major disadvantage of using E. coli host is the production of bodies [30].
Inclusion bodies are insoluble protein aggregates that lack biological function [31]; their formation often occurs when eukaryotic proteins are overexpressed in a heterologous host such as E. coli [32]. Inclusion bodies have been considered a significant obstacle to producing soluble and active recombinant proteins [33][34]. In Table 1, most of the cold-active lipase and esterase were overexpressed in soluble forms, and only five (5) were produced as insoluble or soluble but in inactive forms. It is difficult to explain why most articles examined herein reported more soluble expression than insoluble inclusion bodies. Furthermore, there has been a great success not only in using biochemical and molecular techniques to prevent their formation or to address various challenges during their isolation, solubilisation, refolding, and purification [33], but their biological activity is also emerging [35][36] contrary to the previous notion that they lack activity [31].
Table 1. Cold Active Lipase and Esterase Overexpressed in Heterologous Host.
Organisms/Enzymes Source Host Vector Localization of Expressed Enzyme Optimum Temp./Residual Activity References
Alkalibacterium sp. SL3/esterase Uncultured E. coli BL21 (DE3) pET-28a (+) Soluble 30 °C and 68% at 0 °C [37]
Chitinophaga pinensis-like/esterase Uncultured E. coli RosettaTM (Novagen) pGEX-6P-2 Insoluble inclusion body 20 °C and NA [19]
Lactobacillus plantarum/LpLp_2631/esterase Microbiological Culture E. coli BL21 (DE3) pURI3TEV vector Soluble 20 °C and 90% at 5 °C [38]
Burkholderia pyrrocinia/BpFae esterase Microbiological Culture E. coli BL21 (DE3) pET28a
pCold-TF and pGEX-4T-1.		 Insoluble/soluble non inactive form NA [39]
Candida parapsilosis/esterase Cultured S. cerevisiae pYES2 Soluble NA and at 20 °C [17]
Monascus ruber M7/esterase Cultured E. coli BL21(DE3) pET-30a (+) Soluble 40 °C and 50% at 4–10 °C [40]
Alcanivorax dieselolei/lipase Cultured E. coli BL21(DE3) pGEX-6p-1 (GE) Soluble 20 °C and 95% at 10 °C [41][42]
Pseudomonas fluorescens KE38/lipase Uncultured E. coli BL21(DE3) pET28a Insoluble inclusion body 25 °C and NA [43]
Aphanizomenon flos-aquae/esterase Uncultured E. coli BL21(DE3) pET28a Insoluble inclusion body 5–15 °C [44]
Bacillus halodurans/lipase Uncultured E. coli BL21 (DE3) pET-28a (+) Soluble 30 °C [45]
Bacillus licheniformis/esterase Cultured E. coli BL21 (DE3) pET-28a (+) Soluble 30 °C and 35% at 0 °C [16]
G. antarctica PI12/esterase Expressed sequence tag BL21 (DE3) pET200_GaDlh Soluble 10 °C
and 50% at 0–30 °C		 [46]
Paenibacillus sp. R4/esterase Cultured BL21 (DE3) pET-22b (+) Soluble 35 °C and 45% at 10 °C [47]
Pseudomonas sp./lipase Uncultured BL21(DE3) pET32b (+) Insoluble inclusion body 35 °C and 50% at 15–40 °C [48]
Yarrowia lipolytica(LIPY8)/lipase Cultured Insect (Sf9) pFastBac1 Soluble 17 °C and 70% at 8–30 °C [18]
NA—not available.

3. Purification of Cold-Active Lipolytic Enzymes

Purification is critical in determining an enzyme’s structure and function. Purifying an enzyme not only isolates the target enzyme from other proteins and materials that comprise the crude cell extract but also improves its shelf life and stability. Conformational and structural studies can also be performed after the homogenous purification of the enzymes, and only this homogenous enzyme can be used to establish structure-function relationships [37]. For several decades, protein scientists were into developing screening and optimisation of different combinations of variables during pre-purification and purification experiments Shepard and Tiselius [38] as cited by [39]. The chromatographic pre-purification screening parameters, including resin, ligand, and column screening, are targeted in the experimental design and analytical phases [40]. One example is a high-throughput process development (HTPD) that saves time and cost while harmonising purification procedures through increased automation, miniaturisation, and practical data analysis [41]. A similar format with miniaturised columns enables a high-throughput selection of adsorbent and separation parameters during binding and elution purification experiments. Integrated robot platforms are also employed for choosing a suitable adsorbent in 96-well plates or microcolumn that is essential for determining the success or failure of the purification step [42]. In addition, functionalised microchips, combined with mass spectrometry, are used for protein solution binding, subsequent elution, and analysis. It is possible to determine the optimum binding conditions, the ionic strength for binding, and the lowest ionic strength for the elution [40][43].
Cold-active lipolytic enzymes were purified like other enzymes and proteins sequentially depending on the purity required. For instance, the recommended purity level for structural and functional studies is greater than 98% [44]. Conventional methods include ammonium sulfate precipitation, affinity chromatography, size exclusion (gel filtration), and hydrophobic interaction [37][45][46][47]. Table 2 summarises the various methods used to purify recombinant cold-active lipolytic enzymes. In most cold-active lipase and esterase purification procedures, affinity chromatography is either employed in a one-step or a double-step purification strategy. One-step purification using affinity chromatography generally reduces the time and cost of purification. Even so, the prominent double-step procedure uses ammonium sulfate precipitation with size exclusion and hydrophobic interaction; however, this strategy is suitably employed if the enzymes are produced extracellularly. The affinity chromatography technique is highly specific, while size exclusion, hydrophobic interaction, and ammonium sulphate precipitation are less-specific methods. Sometimes the purpose of using affinity chromatography or ammonium sulphate precipitation in single or first-step purification is to concentrate the recombinant proteins, while less-specific procedures are used to polish the purification. The double-step purification strategy using ammonium sulfate precipitation and nickel affinity has not been utilised much, despite having been reported [49]. In general, obtaining high-purity recombinant enzymes in their stable and active form is expensive, time-consuming, and complex. One-step purification using ammonium sulfate is usually term as partial purification; a well-designed ammonium sulfate precipitation is regarded as a gold standard among several purification strategies [50].
Affinity chromatography is usually achieved by fusing tags at an enzyme’s C or N terminal before its expression [51]. Several affinity tags have been known to facilitate the expression, solubility, detection, and purification of proteins [52][53]. Poly-histidine tagging, also known as His6 or His-tag, is widely employed to express and purify most recombinant proteins, including cold-active lipases and esterase [54]. Despite the high affinity, specificity, and size of His-tag, the technique possesses some disadvantages, including (1) co-purification of other histidine-rich microbial host proteins and (2) negative impact on enzyme stability, activity, binding affinity, and structure [55]. The latter is subject to much contrasting opinion and is still debated because some authors observed that its presence is mainly tolerated for enzymes such as lipase; this cannot be ignored due to its effect on reaction specificity. In a study on the thermal stability of some selected proteins conducted by Booth, Schlachter [56], cleavage of the his-tag can be neutral to some of the proteins while influencing the stability of other protein molecules. In general, the his-tag has an effect (positive or negative) or neutral on proteins.
As shown in Table 2, several scholars have reported a single-step purification of cold-active esterase and lipase using nickel Sepharose or agarose affinity chromatography with good fold and recovery. Furthermore, Noby, Saeed [57] have purified a cold-active esterase EstN7 from Bacillus cohnii strain with 94.5% yield and 5-fold, adopting Tris–HCl (pH 8.0) in the lysis buffer and potassium phosphate (pH 7.5) in the binding buffer differentiate the study from others that utilised the same buffer in both the purification processes. Kim, Park [58], and Lee, Yoo [59] have purified cold-active esterase using a double-step purification that incorporates nickel-affinity and size exclusion chromatography. Another cold-active lipase, B8W22 from Bacillus aryabhattii, was purified in a greater fold of 59.03 using nickel Sepharose affinity and ion-exchange chromatography [60].
Table 2. Purification of Cold-adapted Esterase and Lipase.
Enzymes Type of Purification Purification Steps Buffer Column/Resin Fold/Yield Molecular Mass References
GaDlh Complete Single-step/Ni-affinity chromatography Tris–HCl Ni–NTA column 1.9/7.7% 28 kDa [61]
AMBL-20 Partial Single step/ammonium sulfate precipitation Tris–HCl NA NA NA [62]
HaSGNH1 Complete Single-step/Ni2+-affinity Tris–HCl HisTrap HP 2.5/~5 mg/g 24 kDa [63]
LSK25 Complete Single-step/Ni-Sepharose affinity Tris–HCl Ni Sepharose® 6Fast Flow column 1.3/44% 65 kDa [48]
AaSGNH1 Complete Single-step/Ni-Sepharose affinity Tris–HCl Ni-NTA agarose 0.6–0.7 mg/mL 43.9 kDa [64]
B8W22 Complete Double-step/Ni-Sepharose affinityand ion-exchange Tris–HCl DEAE FF column/Octyl Sepharose FF column 59.03/20% 35 kDa [65]
ERMR1:04 Complete Triple-step/ammonium sulfate precipitation, Size exclusion, and hydrophobic interaction Tris–HCl Sephadex G-100 column, Octyl-Sepharose fast flow column 21.3/NA 250 kDa (hexameric) 39.8 kD (monomeric) [66]
estHIJ Complete Single-step/Ni-affinity Phosphate buffer Ni-NTA affinity column. 3.5/47.5% 29 kDa [67]
ZY124 Complete Double step/ammonium sulfate precipitation and hydrophobic chromatography Tris–HCl Phenyl Sepharose FF column andmicrocolumn reversed-phase LC-1MS 1.34/NA 37.9 kDa. [60]
AMS8 Complete Reverse Micelle Extraction Sodium phosphate NA NA/58.84% NA [68]
KM12 Complete Double-step/ammonium sulfate precipitation and ion-exchange Tris–HCl Q-Sepharose FF column 15.63/36.0% 33 kDa [69]
KCTC 22881 Complete Double-step/affinity chromatography and size-exclusion chromatography Tris–HCl HisTrap FF, PD-10 and Sephacryl S200 HR NA 31.0 kDa [59]
EstN7 Complete Single-step/Ni-affinity Potassium Phosphate Ni–NTA affinity column 5/94.5% 37.0 kDa [57]
GlaEst12-like Complete Single-step/Ni-sepharose affinity Sodium Phosphate Nickel-Sepharose HP 1.7/40% 63 kDa [70]
RSAP17 Complete Double-step/ammonium sulfate precipitation and ion-exchange Tris–HCl DEAE-cellulose anion exchanger NA 103.8 kDa [71]
PsEst3 Complete Double-step/nickel-affinity and size-exclusion chromatography Tris–HCl Ni-affinity and HiLoad 16/60 Superdex 200 column NA 29 kDa [58]
NA—not available.

References

  1. D’Amico, S.; Collins, T.; Marx, J.C.; Feller, G.; Gerday, C. Psychrophilic microorganisms: Challenges for life. EMBO Rep. 2006, 7, 385–389.
  2. Santiago, M.; Ramírez-Sarmiento, C.A.; Zamora, R.A.; Parra, L.P. Discovery, molecular mechanisms, and industrial applications of cold-active enzymes. Front. Microbiol. 2016, 7, 1408.
  3. Gerday, C. Fundamentals of Cold-Active Enzymes. In Cold-adapted Yeasts: Biodiversity, Adaptation Strategies and Biotechnological Significance; Buzzini, P., Margesin, R., Eds.; Springer: Berlin, Heidelberg, 2014; pp. 325–350.
  4. Petrovskaya, L.; Novototskaya-Vlasova, K.; Komolova, A.; Rivkina, E. 6 Biochemical adaptations to the permafrost environment: Lipolytic enzymes from Psychrobacter cryohalolentis K5T. In Microbial Life in the Cryosphere and Its Feedback on Global Change; De Gruyter: Berlin, Germany, 2021; pp. 141–152.
  5. Feller, G. Psychrophilic enzymes: From folding to function and biotechnology. Scientifica 2013, 2013, 512840.
  6. Smalås, A.; Leiros, H.; Os, V.; Willassen, N. Cold adapted enzymes. Biotechnol. Annu. Rev. 2000, 6, 1–57.
  7. Baeza, M.; Alcaíno, J.; Cifuentes, V.; Turchetti, B.; Buzzini, P. Cold-active enzymes from cold-adapted yeasts. In Biotechnology of Yeasts and Filamentous Fungi; Springer: Berlin/Heidelberg, Germany, 2017; pp. 297–324.
  8. Joseph, B.; Ramteke, P.W.; Thomas, G.; Shrivastava, N. Cold-active microbial lipases: A versatile tool for industrial applications. Biotechnol. Mol. Biol. Rev. 2007, 2, 39–48.
  9. Mangiagalli, M.; Brocca, S.; Orlando, M.; Lotti, M. The “cold revolution”. Present and future applications of cold-active enzymes and ice-binding proteins. New Biotechnol. 2020, 55, 5–11.
  10. Kumar, A.; Mukhia, S.; Kumar, R. Industrial applications of cold-adapted enzymes: Challenges, innovations and future perspective. 3 Biotech 2021, 11, 426.
  11. Hamid, B.; Bashir, Z.; Yatoo, A.M.; Mohiddin, F.; Majeed, N.; Bansal, M.; Poczai, P.; Almalki, W.H.; Sayyed, R.; Shati, A.A. Cold-Active Enzymes and Their Potential Industrial Applications—A Review. Molecules 2022, 27, 5885.
  12. Gurung, N.; Ray, S.; Bose, S.; Rai, V. A broader view: Microbial enzymes and their relevance in industries, medicine, and beyond. BioMed Res. Int. 2013, 2013, 329121.
  13. Ramnath, L.; Sithole, B.; Govinden, R. Classification of lipolytic enzymes and their biotechnological applications in the pulping industry. Can. J. Microbiol. 2017, 63, 179–192.
  14. Kleiner-Grote, G.R.M.; Risse, J.M.; Friehs, K. Secretion of recombinant proteins from E. coli. Eng. Life Sci. 2018, 18, 532–550.
  15. Kastenhofer, J.; Rettenbacher, L.; Feuchtenhofer, L.; Mairhofer, J.; Spadiut, O. Inhibition of E. coli host RNA polymerase allows efficient extracellular recombinant protein production by enhancing outer membrane leakiness. Biotechnol. J. 2021, 16, 2000274.
  16. Zhang, W.; Xu, H.; Wu, Y.; Zeng, J.; Guo, Z.; Wang, L.; Shen, C.; Qiao, D.; Cao, Y. A new cold-adapted, alkali-stable and highly salt-tolerant esterase from Bacillus licheniformis. Int. J. Biol. Macromol. 2018, 111, 1183–1193.
  17. Xue, D.; Yao, D.; You, X.; Gong, C. Green Synthesis of the Flavor Esters with a Marine Candida parapsilosis Esterase Expressed in Saccharomyces cerevisiae. Indian J. Microbiol. 2020, 60, 175–181.
  18. Li, T.; Zhang, W.; Hao, J.; Sun, M.; Lin, S.X. Cold-active extracellular lipase: Expression in Sf9 insect cells, purification, and catalysis. Biotechnol. Rep. 2019, 21, e00295.
  19. Hu, X.P.; Heath, C.; Taylor, M.P.; Tuffin, M.; Cowan, D. A novel, extremely alkaliphilic and cold-active esterase from Antarctic desert soil. Extremophiles 2012, 16, 79–86.
  20. Yoshida, K.; Konishi, K.; Magana-Mora, A.; Rougny, A.; Yasutake, Y.; Muramatsu, S.; Murata, S.; Kumagai, T.; Aburatani, S.; Sakasegawa, S.-i. Production of recombinant extracellular cholesterol esterase using consistently active promoters in Burkholderia stabilis. Biosci. Biotechnol. Biochem. 2019, 83, 1974–1984.
  21. Gupta, R.; Gupta, N.; Rathi, P. Bacterial lipases: An overview of production, purification and biochemical properties. Appl. Microbiol. Biotechnol. 2004, 64, 763–781.
  22. Ingram, Z.; Patkar, A.; Oh, D.; Zhang, K.K.; Chung, C.; Lin-Cereghino, J.; LinCereghino, G.P. Overcoming Obstacles in Protein Expression in the Yeast Pichia pastoris: Interviews of Leaders in the Pichia Field. Pac. J. Health 2021, 4, 2.
  23. Karbalaei, M.; Rezaee, S.A.; Farsiani, H. Pichia pastoris: A highly successful expression system for optimal synthesis of heterologous proteins. J. Cell Physiol. 2020, 235, 5867–5881.
  24. Jarvis, D.L. Developing baculovirus-insect cell expression systems for humanized recombinant glycoprotein production. Virology 2003, 310, 1–7.
  25. Wu, C.P.; Chang, C.J.; Li, C.H.; Wu, Y.L. The influence of serial passage on the stability of an exogenous gene expression in recombinant baculovirus. Entomol. Res. 2021, 51, 168–175.
  26. Gomes, A.; Byregowda, S.; Veeregowda, B.; Balamurugan, V. An overview of heterologous expression host systems for the production of recombinant proteins. Adv. Anim. Vet. Sci 2016, 4, 346–356.
  27. Miyauchi, Y.; Kimura, A.; Sawai, M.; Fujimoto, K.; Hirota, Y.; Tanaka, Y.; Takechi, S.; Mackenzie, P.I.; Ishii, Y. Use of a Baculovirus-Mammalian Cell Expression-System for Expression of Drug-Metabolizing Enzymes: Optimization of Infection With a Focus on Cytochrome P450 3A4. Front. Pharmacol. 2022, 13, 832931.
  28. Lozano Terol, G.; Gallego-Jara, J.; Sola Martínez, R.A.; Martínez Vivancos, A.; Cánovas Díaz, M.; de Diego Puente, T. Impact of the Expression System on Recombinant Protein Production in Escherichia coli BL21. Front. Microbiol. 2021, 12, 682001.
  29. Xu, W.; Klumbys, E.; Ang, E.L.; Zhao, H. Emerging molecular biology tools and strategies for engineering natural product biosynthesis. Metab. Eng. Commun. 2020, 10, e00108.
  30. Rosano, G.L.; Ceccarelli, E.A. Recombinant protein expression in Escherichia coli: Advances and challenges. Front. Microbiol. 2014, 5, 172.
  31. Singh, A.; Upadhyay, V.; Upadhyay, A.K.; Singh, S.M.; Panda, A.K. Protein recovery from inclusion bodies of Escherichia coli using mild solubilization process. Microb. Cell Factories 2015, 14, 1–10.
  32. Kane, J.F.; Hartley, D.L. Formation of recombinant protein inclusion bodies in Escherichia coli. Trends Biotechnol. 1988, 6, 95–101.
  33. Slouka, C.; Kopp, J.; Hutwimmer, S.; Strahammer, M.; Strohmer, D.; Eitenberger, E.; Schwaighofer, A.; Herwig, C. Custom made inclusion bodies: Impact of classical process parameters and physiological parameters on inclusion body quality attributes. Microb. Cell Factories 2018, 17, 1–15.
  34. Singhvi, P.; Saneja, A.; Srichandan, S.; Panda, A.K. Bacterial inclusion bodies: A treasure trove of bioactive proteins. Trends Biotechnol. 2020, 38, 474–486.
  35. Ramón, A.; Señorale-Pose, M.; Marín, M. Inclusion bodies: Not that bad…. Front. Microbiol. 2014, 5, 56.
  36. Baltà-Foix, R.; Roca-Pinilla, R.; López-Cano, A.; Gifre-Renom, L.; Arís, A.; Garcia-Fruitós, E. Functional Inclusion Bodies. In Microbial Production of High-Value Products; Rehm, B.H.A., Wibowo, D., Eds.; Springer International Publishing: Cham, Switzerland, 2022; pp. 289–308.
  37. Javed, S.; Azeem, F.; Hussain, S.; Rasul, I.; Siddique, M.H.; Riaz, M.; Afzal, M.; Kouser, A.; Nadeem, H. Bacterial lipases: A review on purification and characterization. Prog. Biophys. Mol. Biol. 2018, 132, 23–34.
  38. Shepard, C.C.; Tiselius, A. The chromatography of proteins. The effect of salt concentration and pH on the adsorption of proteins to silica gel. Discuss. Faraday Soc. 1949, 7, 275–285.
  39. Coffman, J.L.; Kramarczyk, J.F.; Kelley, B.D. High-throughput screening of chromatographic separations: I. Method development and column modeling. Biotechnol. Bioeng. 2008, 100, 605–618.
  40. Bensch, M.; Schulze Wierling, P.; von Lieres, E.; Hubbuch, J. High Throughput Screening of Chromatographic Phases for Rapid Process Development. Chem. Eng. Technol. 2005, 28, 1274–1284.
  41. Silva, T.C.; Eppink, M.; Ottens, M. Automation and miniaturization: Enabling tools for fast, high-throughput process development in integrated continuous biomanufacturing. J. Chem. Technol. Biotechnol. 2021, 97, 2365–2375.
  42. Hollander, C.; Walrond, S.C.; Connelly, C.; Megson, L.; Bove, E.; McDonagh, T. A custom ÄKTA avant configuration enabling automated parallel protein purification over a range of process scales. Protein Expr. Purif. 2021, 182, 105842.
  43. Vorderwuelbecke, S.; Cleverley, S.; Weinberger, S.R.; Wiesner, A. Protein quantification by the SELDI-TOF-MS–based ProteinChip® System. Nat. Methods 2005, 2, 393–395.
  44. Weiss, A.K.H.; Holzknecht, M.; Cappuccio, E.; Dorigatti, I.; Kreidl, K.; Naschberger, A.; Rupp, B.; Gstach, H.; Jansen-Dürr, P. Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins. J. Vis. Exp. 2019, 148, e59729.
  45. Nadeem, H.; Rashid, M.H.; Riaz, M.; Asma, B.; Javed, M.R.; Perveen, R. Invertase from hyper producer strain of Aspergillus niger: Physiochemical properties, thermodynamics and active site residues heat of ionization. Protein Pept. Lett. 2009, 16, 1098–1105.
  46. Kornberg, A. Why purify enzymes? Methods Enzymol. 1990, 182, 1–5.
  47. Ventura, S.P.; Coutinho, J.A. Lipase production and purification from fermentation broth using ionic liquids. In Ionic Liquids in Lipid Processing and Analysis; Elsevier: Amsterdam, The Netherlands, 2016; pp. 59–97.
  48. Salwoom, L.; Salleh, A.B.; Convey, P.; Mohamad Ali, M.S. New recombinant cold-adapted and organic solvent tolerant lipase from psychrophilic Pseudomonas sp. LSK25, isolated from Signy Island Antarctica. Int. J. Mol. Sci. 2019, 20, 1264.
  49. Zhou, X.; Xia, Y. Expression and characterization of recombinant Locusta migratoria manilensis acetylcholinesterase 1 in Pichia pastoris. Protein Expr. Purif. 2011, 77, 62–67.
  50. Koteshwara, A.; Philip, N.V.; Aranjani, J.M.; Hariharapura, R.C.; Volety Mallikarjuna, S. A set of simple methods for detection and extraction of laminarinase. Sci. Rep. 2021, 11, 2489.
  51. Reetz, M.T.; Jaeger, K.-E. Overexpression, immobilization and biotechnological application of Pseudomonas lipases. Chem. Phys. Lipids 1998, 93, 3–14.
  52. Kimple, M.E.; Brill, A.L.; Pasker, R.L. Overview of affinity tags for protein purification. Curr. Protoc. Protein Sci. 2013, 73, 9.9.1–9.9.23.
  53. Fakruddin, M.; Mohammad Mazumdar, R.; Bin Mannan, K.S.; Chowdhury, A.; Hossain, M.N. Critical Factors Affecting the Success of Cloning, Expression, and Mass Production of Enzymes by Recombinant E. coli. ISRN Biotechnol. 2013, 2013, 590587.
  54. de Almeida, J.M.; Moure, V.R.; Müller-Santos, M.; de Souza, E.M.; Pedrosa, F.O.; Mitchell, D.A.; Krieger, N. Tailoring recombinant lipases: Keeping the His-tag favors esterification reactions, removing it favors hydrolysis reactions. Sci. Rep. 2018, 8, 10000.
  55. Meng, L.; Liu, Y.; Yin, X.; Zhou, H.; Wu, J.; Wu, M.; Yang, L. Effects of His-tag on Catalytic Activity and Enantioselectivity of Recombinant Transaminases. Appl. Biochem. Biotechnol. 2020, 190, 880–895.
  56. Booth, W.T.; Schlachter, C.R.; Pote, S.; Ussin, N.; Mank, N.J.; Klapper, V.; Offermann, L.R.; Tang, C.; Hurlburt, B.K.; Chruszcz, M. Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability. ACS Omega 2018, 3, 760–768.
  57. Noby, N.; Saeed, H.; Embaby, A.M.; Pavlidis, I.V.; Hussein, A. Cloning, expression and characterization of cold active esterase (EstN7) from Bacillus cohnii strain N1: A novel member of family IV. Int. J. Biol. Macromol. 2018, 120, 1247–1255.
  58. Kim, H.; Park, A.K.; Lee, J.H.; Shin, S.C.; Park, H.; Kim, H.W. PsEst3, a new psychrophilic esterase from the Arctic bacterium Paenibacillus sp. R4: Crystallization and X-ray crystallographic analysis. Acta Crystallogr. Sect. F Struct. Biol. Commun. 2018, 74, 367–372.
  59. Lee, C.W.; Yoo, W.; Park, S.-H.; Le, L.T.H.L.; Jeong, C.-S.; Ryu, B.H.; Shin, S.C.; Kim, H.-W.; Park, H.; Kim, K.K. Structural and functional characterization of a novel cold-active S-formylglutathione hydrolase (Sf SFGH) homolog from Shewanella frigidimarina, a psychrophilic bacterium. Microb. Cell Factories 2019, 18, 1–13.
  60. Zhang, Y.; Ji, F.; Wang, J.; Pu, Z.; Jiang, B.; Bao, Y. Purification and characterization of a novel organic solvent-tolerant and cold-adapted lipase from Psychrobacter sp. ZY124. Extremophiles 2018, 22, 287–300.
  61. Hashim, N.H.F.; Mahadi, N.M.; Illias, R.M.; Feroz, S.R.; Abu Bakar, F.D.; Murad, A.M.A. Biochemical and structural characterization of a novel cold-active esterase-like protein from the psychrophilic yeast Glaciozyma antarctica. Extremophiles 2018, 22, 607–616.
  62. Yasin, M.T.; Ali, Y.; Ahmad, K.; Ghani, A.; Amanat, K.; Basheir, M.M.; Faheem, M.; Hussain, S.; Ahmad, B.; Hussain, A.; et al. Alkaline lipase production by novel meso-tolerant psychrophilic Exiguobacterium sp. strain (AMBL-20) isolated from glacier of northeastern Pakistan. Arch. Microbiol. 2021, 203, 1309–1320.
  63. Le, L.T.H.L.; Yoo, W.; Jeon, S.; Lee, C.; Kim, K.K.; Lee, J.H.; Kim, T.D. Biodiesel and flavor compound production using a novel promiscuous cold-adapted SGNH-type lipase (HaSGNH1) from the psychrophilic bacterium Halocynthiibacter arcticus. Biotechnol. Biofuels 2020, 13, 55.
  64. Knepp, Z.J.; Ghaner, A.; Root, K.T. Purification and refolding protocol for cold-active recombinant esterase AaSGNH1 from Aphanizomenon flos-aquae expressed as insoluble inclusion bodies. Prep. Biochem. Biotechnol. 2021, 52, 394–403.
  65. Zhang, Y.-J.; Chen, C.-S.; Liu, H.-T.; Chen, J.-L.; Xia, Y.; Wu, S.-J. Purification, identification and characterization of an esterase with high enantioselectivity to (S)-ethyl indoline-2-carboxylate. Biotechnol. Lett. 2019, 41, 1223–1232.
  66. Kumar, A.; Mukhia, S.; Kumar, N.; Acharya, V.; Kumar, S.; Kumar, R. A Broad Temperature Active Lipase Purified From a Psychrotrophic Bacterium of Sikkim Himalaya With Potential Application in Detergent Formulation. Front. Bioeng. Biotechnol. 2020, 8, 642.
  67. Noby, N.; Hussein, A.; Saeed, H.; Embaby, A.M. Recombinant cold-adapted halotolerant, organic solvent-stable esterase (estHIJ) from Bacillus halodurans. Anal. Biochem. 2020, 591, 113554.
  68. Salleh, A.B.; Mohamad Ali, M.S. Optimization and in silico analysis of a cold-adapted lipase from an antarctic Pseudomonas sp. strain ams8 reaction in triton x-100 reverse micelles. Catalysts 2018, 8, 289.
  69. Malekabadi, S.; Badoei-dalfard, A.; Karami, Z. Biochemical characterization of a novel cold-active, halophilic and organic solvent-tolerant lipase from B. licheniformis KM12 with potential application for biodiesel production. Int. J. Biol. Macromol. 2018, 109, 389–398.
  70. Mohamad Tahir, H.; Raja Abd Rahman, R.N.Z.; Chor Leow, A.T.; Mohamad Ali, M.S. Expression, Characterisation and Homology Modelling of a Novel Hormone-Sensitive Lipase (HSL)-Like Esterase from Glaciozyma antarctica. Catalysts 2020, 10, 58.
  71. Uddin, M.R.; Roy, P.; Mandal, S. Production of extracellular lipase from psychrotrophic bacterium Oceanisphaera sp. RSAP17 isolated from arctic soil. Antonie Leeuwenhoek 2021, 114, 2175–2188.
More
© Text is available under the terms and conditions of the Creative Commons Attribution (CC BY) license; additional terms may apply. By using this site, you agree to the Terms and Conditions and Privacy Policy.
Upload a video for this entry
Information
Subjects: Biochemistry & Molecular Biology
Contributors MDPI registered users' name will be linked to their SciProfiles pages. To register with us, please refer to https://encyclopedia.pub/register : Adamu Idris Matinja , Nor Hafizah Ahmad Kamarudin , Adam Thean Chor Leow , Siti Nurbaya Oslan , Mohd Shukuri Mohamad Ali
View Times: 907
Revisions: 2 times (View History)
Update Date: 06 Jan 2023
Table of Contents
    Notice
    You are not a member of the advisory board for this topic. If you want to update advisory board member profile, please contact office@encyclopedia.pub.
    OK
    Confirm
    Only members of the Encyclopedia advisory board for this topic are allowed to note entries. Would you like to become an advisory board member of the Encyclopedia?
    Yes
    No
    ${ textCharacter }/${ maxCharacter }
    Submit
    Cancel
    There is no comment~
    ${ textCharacter }/${ maxCharacter }
    Submit
    Cancel
    ${ selectedItem.replyTextCharacter }/${ selectedItem.replyMaxCharacter }
    Submit
    Cancel
    Confirm
    Are you sure to Delete?
    Yes No
    Academic Video Service
    Feedback