Immobilization of Yarrowia lipolytica Lipase—a Comparison of Stability of Physical Adsorption and Covalent Attachment Techniques

Abstract

Lipase immobilization offers unique advantages in terms of better process control, enhanced stability, predictable decay rates and improved economics. This work evaluated the immobilization of a highly active Yarrowia lipolytica lipase (YLL) by physical adsorption and covalent attachment. The enzyme was adsorbed on octyl-agarose and octadecyl-sepabeads supports by hydrophobic adsorption at low ionic strength and on MANAE-agarose support by ionic adsorption. CNBr-agarose was used as support for the covalent attachment immobilization. Immobilization yields of 71, 90 and 97% were obtained when Y. lipolytica lipase was immobilized into octyl-agarose, octadecyl-sepabeads and MANAE-agarose, respectively. However, the activity retention was lower (34% for octyl-agarose, 50% for octadecyl-sepabeads and 61% for MANAE-agarose), indicating that the immobilized lipase lost activity during immobilization procedures. Furthermore, immobilization by covalent attachment led to complete enzyme inactivation. Thermal deactivation was studied at a temperature range from 25 to 45 degrees C and pH varying from 5.0 to 9.0 and revealed that the hydrophobic adsorption on octadecyl-sepabeads produced an appreciable stabilization of the biocatalyst. The octadecyl-sepabeads biocatalyst was almost tenfold more stable than free lipase, and its thermal deactivation profile was also modified. On the other hand, the Y. lipolytica lipase immobilized on octyl-agarose and MANAE-agarose supports presented low stability, even less than the free enzyme.

Immobilization of a Commercial Lipase from Penicillium camembertii (Lipase G) by Different Strategies

“…The first methodology to be tested was the physical adsorption on highly hydrophobic supports with different characteristics such as octyl-agarose, PHB, and amberlite XAD-4 (Table 1) to take advantage of the complex mechanism of lipases as a tool that allows the immobilization via hydrophobic interaction at very low ionic strength [9, 20, 21]. Lipases recognize these surfaces similarly to those of their natural substrates (drops of oil), yielding immobilized derivative with open and hyperactivated structures [9, 21].…”

“…These enzymes are strongly adsorbed onto hydrophobic interfaces through the lid that covers their active site, recognizing these supports as their natural substrate (hydrophobic oil interfaces). This technique is easy, cheap, and allows facile recycling of the support at the end of the life of the enzyme [9, 20–22]. Enzymes can be also reversibly immobilized on ionic exchange supports.…”

Biodiesel production using cross-linked Staphylococcus haemolyticus lipase immobilized on solid polymeric carriers

“…Instead, lipase enzymes could be applied for biodiesel production in an eco-friendly manner [9,11]. For decades, much of the literature relevant to this topic has been focused largely on the process of biodiesel production using microbial lipases [9,12]. Among these, Candida antarctica lipase B (CalB) has been most frequently utilized, owing primarily to its profound ability to carry out the biodiesel production reaction with a variety of plant oils.…”

“…Many diverse immobilization methods have been developed for this purpose. Among them are some methods which require rigid supports: most notably, hydrophobic adsorption [12], ionic immobilization [15,16], and covalent immobilization [17][18][19]. Choosing the most suitable immobilization method has become an important factor for transesterification efficiency of microbial lipases.…”

Comparison of the Properties of Lipase Immobilized onto Mesoporous Resins by Different Methods

“…The treatment of adsorbed enzyme on support with cross-linker can solve the above problems to a certain extent [9][10][11][12][13]. Glutaraldehyde is the most effective cross-linker [14][15][16], which can enhance the interactions between enzyme and support.…”