Crosslinked enzyme aggregates (CLEA) of phytase with soymilk proteins

Harini, Tirunagari; Basetty, Shalini; Rode, Haridas B.; Fadnavis, Nitin W.

doi:10.1016/j.jbiotec.2018.07.003

Cited by 22 publications

(8 citation statements)

References 32 publications

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“…This may be determined by measuring the activity of the suspension and supernatants in each washing step. The solution may be to add media rich in primary amino groups, proteins or polymers, amination of the enzyme or alternatively use other cross-linkers (159,160,161,162,163,164,165,166,167,168).…”

Section: There Is Not An Actual Immobilization Time Coursementioning

confidence: 99%

Parameters necessary to define an immobilized enzyme preparation

Boudrant

Woodley

Fernández-Lafuente

2020

Process Biochemistry

354

177

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Biocatalytic processes continue to find increasing application in industry. Therefore enzyme immobilization has also become of increasing importance as a means of allowing enzyme containment within reactors operating in continuous mode or else separation of enzyme after use in (fed-)batch reactors, as well as potential recycle. Whilst much has been reported in the scientific literature about enzyme immobilization methods, in many cases the protocol leads to losses in enzyme activity. In this review we outline the reasons for loss of activity during immobilization and highlight suitable diagnostic tests to elucidate the precise cause and thereby methods to restore activity. The need for standardized reporting of immobilization methods is also emphasized as a means of benchmarking alternative approaches.

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Section: There Is Not An Actual Immobilization Time Coursementioning

confidence: 99%

Parameters necessary to define an immobilized enzyme preparation

Boudrant

Woodley

Fernández-Lafuente

2020

Process Biochemistry

354

177

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“…[40][41][42][43] A company (CLEA technology) was created to commercialize CLEAs from different enzymes. However, the poor mechanical resistance makes that this biocatalysts must be utilized in reactors that do not submit the particles to mechanical stress, such as basket [44][45][46] or vortex reactors. 47 In fact, some researchers have proposed the trapping of the cross-linked aggregates in solids with better mechanical features [48][49][50][51] Other very popular strategy of enzyme immobilization in ex novo solids is the formation of nano-flowers, where the enzyme is incubated in some metal salts and a metal crystal grows using the enzyme surface as nucleation points.…”

Section: Introductionmentioning

confidence: 99%

Is enzyme immobilization a mature discipline? Some critical considerations to capitalize on the benefits of immobilization

2022

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“…The treatment with PEI was performed according to Wilson et al [ 49 ], adding 50 μL of PEI solution (100 mg mL −1 ) in a homogeneous PPL solution (5 mg mL −1 ) prepared in 5 mM phosphate buffer (final pH 7.0). The reaction medium was incubated at 25 °C and stirred at 150 rpm for 60 min.…”

Section: Methodsmentioning

confidence: 99%

“…When the enzyme surface has a low content of amino groups, co-feeders are commonly used to aid the crosslinking, such as proteins (bovine serum albumin, soy protein, etc.) [ 15 , 45 , 46 , 47 , 48 , 49 ], polyethyleneimine [ 50 , 51 ], and so on. CLEAs of several enzymes have been reported [ 12 , 15 , 47 , 52 , 53 , 54 , 55 ], including bovine and porcine pancreas lipases.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Strategies to Produce Highly Porous Cross-Linked Aggregates of Porcine Pancreas Lipase with Magnetic Properties

et al. 2018

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The preparation of highly porous magnetic crosslinked aggregates (pm-CLEA) of porcine pancreas lipase (PPL) is reported. Some strategies to improve the volumetric activity of the immobilized biocatalyst were evaluated, such as treatment of PPL with enzyme surface-modifying agents (polyethyleneimine or dodecyl aldehyde), co-aggregation with protein co-feeders (bovine serum albumin and/or soy protein), use of silica magnetic nanoparticles functionalized with amino groups (SMNPs) as separation aid, and starch as pore-making agent. The combination of enzyme surface modification with dodecyl aldehyde, co-aggregation with SMNPs and soy protein, in the presence of 0.8% starch (followed by hydrolysis of the starch with α-amylase), yielded CLEAs expressing high activity (immobilization yield around 100% and recovered activity around 80%), high effectiveness factor (approximately 65% of the equivalent free enzyme activity) and high stability at 40 °C and pH 8.0, i.e., PPL CLEAs co-aggregated with SMNPs/bovine serum albumin or SMNPs/soy protein retained 80% and 50% activity after 10 h incubation, respectively, while free PPL was fully inactivated after 2 h. Besides, highly porous magnetic CLEAs co-aggregated with soy protein and magnetic nanoparticles (pm-SP-CLEAs) showed good performance and reusability in the hydrolysis of tributyrin for five 4h-batches.

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Crosslinked enzyme aggregates (CLEA) of phytase with soymilk proteins

Cited by 22 publications

References 32 publications

Parameters necessary to define an immobilized enzyme preparation

Parameters necessary to define an immobilized enzyme preparation

Is enzyme immobilization a mature discipline? Some critical considerations to capitalize on the benefits of immobilization

Evaluation of Strategies to Produce Highly Porous Cross-Linked Aggregates of Porcine Pancreas Lipase with Magnetic Properties

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