A highly xyloglucan active lytic polysaccharide monooxygenase EpLPMO9A from Eupenicillium parvum 4-14 shows boosting effect on hydrolysis of complex lignocellulosic substrates

Shi, Yuexin; Chen, Kaixiang; Long, Liangkun; Ding, Shaojun

doi:10.1016/j.ijbiomac.2020.11.177

Cited by 12 publications

(5 citation statements)

References 56 publications

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“…The HPLC system tted with a Bio-Rad Aminex HPX-87H column was operated at 55 °C with 5 mM H 2 SO 4 as the mobile phase at the ow rate of 0.6 mL/min as previously described by Chen et al (65). HPAEC-PAD analysis was performed on a Dionex ICS-5000 system (Dionex, Sunnyvale, CA, USA) equipped with pulsed amperometric detection (PAD) and a CarboPac PA200 analytical column (3 × 250 mm) with a CarboPac PA200 guard column (3 × 50 mm) according to Shi et al described method (13). MALDI-TOF MS analysis was performed on a 5800 MALDI TOF/TOF analyzer (AB SCIEX, Foster City, CA, USA) equipped with a neodymium: yttrium-aluminum-garnet laser (laser wavelength was 349 nm) according to Shi et al described method (13).…”

Section: Test Methodsmentioning

confidence: 99%

“…The hydrolytic e ciency and cost of enzymes in the second step are major factors restricting the cost-effective production of biofuels and biochemicals from lignocellulose due to biomass recalcitrance (11). In view of the complex and strong network structure formed by XG with other lignocellulosic components, XG is very di cult to be removed during common pretreatment, and may be still remained in the pretreated lignocellulosic biomasses, therefore limiting the accessibility and hydrolysis e ciency of cellulase to cellulose (12,13). The removal of XG is needed to promote the enzymatic hydrolysis of pretreated lignocellulose.…”

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confidence: 99%

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Comparison of a GH74 xyloglucanase and its CBM-deleted variant from Thielavia terrestris and their roles in xyloglucan-rich biomass degradation

Wang

Chen

Zhang

et al. 2022

Preprint

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Background The hydrolysis of lignocellulose was greatly hindered by the associated hemicellulose. Xyloglucan (XG) is a major complex hemicellulose in the primary plant cell wall, however, the blocking effect of xyloglucan on lignocellulose biodegradation was still less understood compared with xylan and mannan. Glycoside hydrolase family 74 (GH74) enzymes are specific xyloglucanases widely distributed in bacteria and fungi. Fungal GH74 xyloglucanases generally contains family 1 carbohydrate binding module (CBM1), but its effect on the enzymatic properties of GH74 xyloglucanases and its roles in xyloglucan-rich biomass degradation has not been investigated in depth. Results TtGH74 and CBM-deleted variant (TtGH74ΔCBM) from Thielavia terrestris had the same optimum temperature and pH, but TtGH74ΔCBM had higher thermostability. TtGH74 displayed high binding affinity on xyloglucan and cellulose, while TtGH74ΔCBM almost completely lost the adsorption capacity on cellulose. Their hydrolysis action alone or combined with other glycoside hydrolases on the free XG, xyloglucan-coated phosphoric acid swollen cellulose or pretreated corn bran and apple pomace was compared in this study. CBM1 might not be essential for hydrolysis of free XG, but effective for the associated XG to some extent. TtGH74 was more effective in hydrolysis of xyloglucan in corn bran, while TtGH74ΔCBM showed relatively higher catalytic activity on apple pomace. Similar phenomenon was also observed when TtGH74 or TtGH74ΔCBM synergistically acting with CBH1/EG1 mixture. The addition of TtGH74 or TtGH74ΔCBM in CBH1/EG1 mixture significantly facilitated the overall hydrolysis of pretreated corn bran and apple pomace by 1.22–2.02 folds in terms of the degree of synergy. The extra addition of GH10 xylanase in TtGH74 or TtGH74ΔCBM /CBH1/EG1 mixture further improved the overall hydrolysis efficiency, and the degree of synergy was up to 1.50–2.16 because of co-existence of xyloglucan and xylan in pretreated corn bran and apple pomace. Conclusions TtGH74 can remove the bound xyloglucan on cellulose, therefore efficiently boost the hydrolysis of the pretreated lignocellulosic biomass by synergistic action with cellulase and xylanase. The presence of CBM1 in TtGH74 is conducive to enzymatic hydrolysis, but its role and significance is substrate-specific due to the differences of xyloglucan contents and structure in various biomasses.

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Section: Test Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Comparison of a GH74 xyloglucanase and its CBM-deleted variant from Thielavia terrestris and their roles in xyloglucan-rich biomass degradation

Wang

Chen

Zhang

et al. 2022

Preprint

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“…The reaction was stopped by boiling at 99 °C for 10 min and centrifuging at 10,000 rpm for 10 min. The products in the supernatant were detected by HPAEC-PAD as described previously, 24 and the sum of C4-oxidized product peak areas (20−22 min) was calculated using Chameleon 6 software (Dionex).…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

“…The transient electron-transfer kinetics was studied by using stopped-flow spectroscopy. Besides, the IPET between the two CDHs and AA9 LPMOs (C4-oxidizing Nc LPMO9C and C1/C4-oxidizing Ep LPMO9A from Eupenicillium parvum 4–14) and the efficiencies in driving the reaction of two AA9 LPMOs under different pH and divalent cation (Ca 2+ and Mn 2+ ) conditions were also investigated. We demonstrated that the unique lysine-rich sequence on the CYT domain of Af CDH played a very important role in the interaction between DH with CYT and significantly enhanced the IDET rate in CDH and the reductive activation of LPMOs.…”

Section: Introductionmentioning

confidence: 99%

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Unique Lysine-Rich Sequence on the CYT Domain of AfCDH Enhances Its Interdomain Electron Transfer and Activation of AA9 LPMOs

Chen

Yang

Ding

et al. 2022

ACS Sustainable Chem. Eng.

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Cellobiose dehydrogenase (CDH) is a biologically relevant electron donor for lytic polysaccharide monooxygenase (LPMO) in the oxidative cleavage of recalcitrant polysaccharides, so exploring the intrinsic factors affecting the electron transfer from CDH to LPMO is vital for governing the CDH-driven LPMO reaction efficiency. We previously found a unique lysine-rich sequence on the cytochrome (CYT) domain of Af CDH from Aspergillus fumigatus. Here, we comprehensively compared the functionality of Af CDH and Af CDHΔ (the lysine-rich sequencedeleted mutant) to elucidate the role and significance of the lysinerich sequence in its interdomain electron transfer (IDET) and activation of LPMOs. The results showed that the lysine-rich sequence had no influence on the dehydrogenase (DH) reductive half-reaction and the interprotein electron transfer (IPET) rate from CYT to LPMOs. However, a remarkably faster IDET rate between DH and CYT was observed in Af CDH compared to that in Af CDHΔ, especially under high pH conditions. Moreover, Af CDH was found to be more effective in driving the LPMO reactions than Af CDHΔ. Our findings demonstrated that IDET is crucial for the overall electron transfer from CDH to LPMO and the CDHdriven LPMO reaction efficiency. This work provides a feasible strategy for engineering the lysine-rich sequence on CYT to potentiate the IDET rate in other CDHs and for the activation of LPMO.

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Two C1-oxidizing AA9 lytic polysaccharide monooxygenases from Sordaria brevicollis differ in thermostability, activity, and synergy with cellulase

Zhang

Chen

Long

et al. 2021

Appl Microbiol Biotechnol

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A highly xyloglucan active lytic polysaccharide monooxygenase EpLPMO9A from Eupenicillium parvum 4-14 shows boosting effect on hydrolysis of complex lignocellulosic substrates

Cited by 12 publications

References 56 publications

Comparison of a GH74 xyloglucanase and its CBM-deleted variant from Thielavia terrestris and their roles in xyloglucan-rich biomass degradation

Comparison of a GH74 xyloglucanase and its CBM-deleted variant from Thielavia terrestris and their roles in xyloglucan-rich biomass degradation

Unique Lysine-Rich Sequence on the CYT Domain of AfCDH Enhances Its Interdomain Electron Transfer and Activation of AA9 LPMOs

Two C1-oxidizing AA9 lytic polysaccharide monooxygenases from Sordaria brevicollis differ in thermostability, activity, and synergy with cellulase

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