Temperature and pH influence adsorption of cellobiohydrolase onto lignin by changing the protein properties

Lu, Xianqin; Wang, Can; Li, Xuezhi; Zhao, Jian

doi:10.1016/j.biortech.2017.08.139

Cited by 42 publications

(19 citation statements)

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“…According to our previous report, elevating the pH value can reduce the adsorption of cellulase to lignin by changing the protein surface properties and by enhancing the repulsion between lignin and enzyme, thus leading to an improvement in the hydrolysis yield [28]. As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

Enzymatic sugar production from elephant grass and reed straw through pretreatments and hydrolysis with addition of thioredoxin-His-S

Zhang

et al. 2019

Biotechnol Biofuels

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BackgroundThe bioconversion of lignocellulose to fermentable C5/C6-saccharides is composed of pretreatment and enzymatic hydrolysis. Lignin, as one of the main components, resists lignocellulose to be bio-digested. Alkali and organosolv treatments were reported to be able to delignify feedstocks and loose lignocellulose structure. In addition, the use of additives was an alternative way to block lignin and reduce the binding of cellulases to lignin during hydrolysis. However, the relatively high cost of these additives limits their commercial application.ResultsThis study explored the feasibility of using elephant grass (Pennisetum purpureum) and reed straw (Phragmites australis), both of which are important fibrous plants with high biomass, no-occupation of cultivated land, and soil phytoremediation, as feedstocks for bio-saccharification. Compared with typical agricultural residues, elephant grass and reed straw contained high contents of cellulose and hemicellulose. However, lignin droplets on the surface of elephant grass and the high lignin content in reed straw limited their hydrolysis performances. High hydrolysis yield was obtained for reed straw after organosolv and alkali pretreatments via increasing cellulose content and removing lignin. However, the hydrolysis of elephant grass was only enhanced by organosolv pretreatment. Further study showed that the addition of bovine serum albumin (BSA) or thioredoxin with His- and S-Tags (Trx-His-S) improved the hydrolysis of alkali-pretreated elephant grass. In particular, Trx-His-S was first used as an additive in lignocellulose saccharification. Its structural and catalytic properties were supposed to be beneficial for enzymatic hydrolysis.ConclusionsElephant grass and reed straw could be used as feedstocks for bioconversion. Organosolv and alkali pretreatments improved their enzymatic sugar production; however, the increase in hydrolysis yield of pretreated elephant grass was not as effective as that of reed straw. During the hydrolysis of alkali-pretreated elephant grass, Trx-His-S performed well as additive, and its structural and catalytic capability was beneficial for enzymatic hydrolysis.

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Section: Resultsmentioning

confidence: 99%

Enzymatic sugar production from elephant grass and reed straw through pretreatments and hydrolysis with addition of thioredoxin-His-S

Zhang

et al. 2019

Biotechnol Biofuels

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“…Subsequently, the adsorption of cellulase onto lignins has slightly reduction. This probably because the adsorption of cellulase onto lignin effected by the different adsorption behavior of monocomponent of cellulase onto lignin, which needs further investigation (Li et al 2017;Kellock et al 2017 ). From the results shown in Fig.…”

Section: Table 3 Langmuir Adsorption Isotherm Parameters Of Enzyme Amentioning

confidence: 98%

“…Other than hydrophobicity, the surface charge of lignin, and the stability of cellulase have been found to synergistically influence the cellulase-lignin interaction (Lu et al 2017). Different types of enzyme may differ in their interaction with lignin.…”

Section: Adsorption Capacity Of Different Enzymes Onto the Ligninmentioning

confidence: 99%

“…Kellock et al (2017) indicated that lignins isolated from steam pretreated spruce (SPS) is more inhibitory to xylanase, β-glucosidase, and individual components of cellulase such as CBH and endoglucanases (EG) than wheat straw (SPWS) lignin. The pH value and temperature could influence the adsorption and desorption behaviors of cellulase on lignin by changing the protein properties (Lu et al 2017). It was shown that enzyme properties, surface charge, and surface hydrophobicity could not alone explain the adsorption behaviour of enzyme onto lignins (Kellock et al 2017).…”

Section: Introductionmentioning

confidence: 99%

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Study of the Difference between Enzyme Adsorption onto Hydrotropic and Alkali Lignin Separated from Eucalyptus and Bamboo

Mou¹,

Wu²,

He³

et al. 2018

BioResources

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Enzymatic hydrolysis of lignocellulosic biomass is the key step for controlling the cost of bioethanol production. However, the non-productive adsorption of cellulase onto lignin in biomass severely hampers the enzyme activity and hydrolysis efficiency. Thus, understanding the adsorption mechanism of cellulase onto lignin is critical for the development of enzyme mixtures and enzymatic hydrolysis. In this investigation, cellulase, β-glucosidase (BG), and xylanase adsorption onto lignin from eucalyptus and bamboo, extracted by alkali and hydrotropic techniques, were compared. The physico-chemical properties of the four types of isolated lignin were detected. Langmuir isotherms were used to interpret the cellulase adsorption kinetics of the lignin. The hydrophobicity was found to be the major factor that affected the cellulase adsorption affinity of lignin. The surface charge was important for the adsorption of BG and xylanase onto the lignin. A comparison was made between hydrotropic and alkali lignin, and the hydrotropic lignin from eucalyptus had the highest cellulase adsorption capacity and lowest BG and xylanase adsorption capacities.

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“…The non-productive adsorption of cellulases onto lignin in biomass is a key economic issue for biofuel processing, since higher cellulase loadings are required to overcome the inhibitory effect of lignin, and cellulase recycling is made difficult after the hydrolysis reaction [9–13]. Many cellulases are multi-domain enzymes consisting of a catalytic domain (CD) and a carbohydrate-binding module (CBM) connected by a flexible linker peptide.…”

Section: Introductionmentioning

confidence: 99%

Altering the linker in processive GH5 endoglucanase 1 modulates lignin binding and catalytic properties

Wang

Zhang

Long

et al. 2018

Biotechnol Biofuels

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BackgroundThe non-productive adsorption of cellulases onto lignin in biomass is a key issue for the biofuel process economy. It would be helpful to reduce the inhibitory effect of lignin on enzymatic hydrolysis by engineering weak lignin-binding cellulases. Cellulase linkers are highly divergent in their lengths, compositions, and glycosylations. Numerous studies have revealed that linkers can facilitate optimal interactions between structured domains. Recently, efforts have focused on the contributions and mechanisms of carbohydrate-binding modules and catalytic domains that affect lignin affinity and processivity of cellulases, but our understanding of the effects of the linker regions on lignin adsorption and processivity of GH5 processive endoglucanases is still limited.ResultsEight GH5 endoglucanase 1 variants of varying length, flexibility, and sequence in the linker region were constructed. Their characteristics were then compared to the wild-type enzyme (EG1). Remarkably, significant differences in the lignin adsorption profiles and processivities were observed for EG1 and other variants. Our studies suggest that either the length or the specific amino acid composition of the linker has a prominent influence on the lignin-binding affinity of the enzymes. Comparatively, the processivity may depend primarily on the length of the linker and less so on the specific amino acid composition. EG1-ApCel5A, a variant with better performance in enzymatic hydrolysis in the presence of lignin, was obtained by replacing a longer, flexible linker. In total, up to between 28.2 and 30.1% more reducing sugars were generated from filter paper by EG1-ApCel5A in the presence of lignin compared to EG1.ConclusionsOur results highlight the relevance of the linker region in the lignin adsorption and processivity of a processive endoglucanase. Our findings suggest that the linker region may be used as a target for the design of more active and weaker lignin-binding cellulases.Electronic supplementary materialThe online version of this article (10.1186/s13068-018-1333-3) contains supplementary material, which is available to authorized users.

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Temperature and pH influence adsorption of cellobiohydrolase onto lignin by changing the protein properties

Cited by 42 publications

References 41 publications

Enzymatic sugar production from elephant grass and reed straw through pretreatments and hydrolysis with addition of thioredoxin-His-S

Enzymatic sugar production from elephant grass and reed straw through pretreatments and hydrolysis with addition of thioredoxin-His-S

Study of the Difference between Enzyme Adsorption onto Hydrotropic and Alkali Lignin Separated from Eucalyptus and Bamboo

Altering the linker in processive GH5 endoglucanase 1 modulates lignin binding and catalytic properties

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