Advances in the Mechanism of Action, Characteristics, and

Applications of Alginate Lyases

Guanchu Ma

1,a

, Chen Zhao

3,4,b

, Kunlun Li

2,c

, Le Su

1,d

, Song Zhang

1,e

, Qiulin Yue

1,f,†

and Lin Zhao

1,g,†

State Key Laboratory of Biobased Material and Green Papermaking, Shandong Provincial Key Laboratory of Microbial

Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250353, China

Jinan Hangchen Biotechnology Co., Ltd., Jinan 250353, China

National Engineering Laboratory of Crop Stress Resistance Breeding, Anhui Agricultural University, Hefei 230036,

People’s Republic of China

Shandong Food Ferment Industry Research & Design Institute, Qilu University of Technology (Shandong Academy of

Sciences), Jinan 250353, P.R. China

f,†

yueqiulin88@163.com,

g,†

sdilizhaolin@163.com

Keywords: Alginate Lyase, Mechanism of Action, Enzyme Characterization, Application.

Abstract: Alginate is a natural polysaccharide. Alginate lyase could break the glycosidic bonds by β-elimination and

form the unsaturated double bonds to complete the degradation of alginate. With the development of enzyme

resources, screening and extracting valuable enzymes from various organisms and developing polysaccharide

degradation products have become an important aspect of the development of biological resources.

Combining various researches, this paper summarizes the research progress of alginate lyase in recent years

from the mechanism of action mechanism, enzyme characterizations and enzyme application.

1 INTRODUCTION

Alginate is mainly a by-product of iodine and

mannitol extracted from marine algae, which mainly

exists in cell wall and intracellular substances. It is a

copolymer composed of α-L- guluronic and its C5

isomer β-D-mannuronic in different proportions

through β-1,4-glycosidic bond. At present, alginate

oligosaccharides are used in various work and

research, such as plant immune (Gurpilhares, 2019),

food storage (Aitouguinane, 2020), biomedicine

(Zhao, 2020), biofuel (Lu, 2019) and other fields.

There are three main degradation methods of

alginate: chemical method, physical method and

enzymatic hydrolysis. In comparison, the advantage

of enzymatic degradation is more obvious, its

efficiency is high, the reaction conditions are mild,

the process is easy to control and has little impact on

the environment, so it is a better method to prepare

alginate oligosaccharides. Therefore, in recent years,

the research on alginate lyases, one of the seaweed

tool enzymes and its degradation product, alginate

oligosaccharides, has attracted increasing attention.

It is found that alginate lyases have important

commercial application value for the development of

bioengineering products. It can not only develop new

physiologically active substances, but more

importantly, it opens up a new field of industrial

application. A more important application trend is to

focus on the enzymatic production of algal

oligosaccharides, providing new means for the

research and development of biomedical production

and food technology, especially the physiological

effects of alginate oligosaccharides, an enzymatic

hydrolysate called oligosaccharide, have been

revealed, which has aroused more interest.

2 MECHANISM OF ACTION AND

ENZYME

CHARACTERIZATION

Alginate lyases catalyze the degradation of alginate

through β-elimination, an unsaturated double bond

was formed between C4 and C5, and a non-reducing

end 4-deoxy-β-L-erythro-hex-4-

enopyranosyluronicacid was generated, the conjugate

of unsaturated oligosaccharides in double bonds and

carboxyl groups results in a strong absorption peak at

Ma, G., Zhao, C., Li, K., Su, L., Zhang, S., Yue, Q. and Zhao, L.

Advances in the Mechanism of Action, Characteristics, and Applications of Alginate Lyases.

DOI: 10.5220/0011181800003443

In Proceedings of the 4th International Conference on Biomedical Engineering and Bioinformatics (ICBEB 2022), pages 75-78

ISBN: 978-989-758-595-1

235nm for oligosaccharide products (Natsume,

1994). Gacesa et al. have hypothesized that the

process of enzymatic hydrolysis of alginate by

alginate lyase can be divided into three steps, namely:

(a) eliminate the negative charge on the carboxyl ion

by salt bridge neutralization reaction; (b) attract

protons on the C5 position; (c) The unsaturated

double bond between C4 and C5 is formed by β-

elimination of the transfer of electrons from the

carboxyl group (Gacesa, 1987).

Recently, a large number of scientific

achievements of alginate lyase have been reported by

researchers, and their enzymatic properties have also

been studied through heterologous expression. It has

filled the blank of alginate lyases research, and

provided sufficient experimental verification for

future commercial application (Table 1). The

enzymatic properties of alginate lyases produced by

different species have certain differences. Most

enzymes have milder working conditions, the

optimum temperature is between 30-50℃, the

optimum pH is 7.0-8.0, neutral and alkaline. But there

are exceptions. For example, in 2016, InoueA et al.,

the alginate lyase NitAly produced by the strain

Nitratiruptor sp.SB155-2 isolated interestingly, its

optimal temperature could reach 70°C (Inoue, 2016).

Metal ions and their concentrations also have an

important impact on the enzymatic activity of

alginate lyases. Wang et al. reported in a 2019 study

that for Vibrio sp. SY01, its alginate lyase Aly08

produced by it will enhance its activity in an

environment containing NaCl, and will reach a peak

activity at 300 mM, which is about more than 8 times

in the non-NaCl environment. In the presence of other

metal ions such as NH

, Li

, Zn

, Ba

and Co

Aly08 has no obvious activation effect. Because of

and Mn

, its enzyme activity increases greatly,

while the surfactants SDS and EDTA seriously

weakened the activity of alginate lyase, and the

relative activity decreased by more than 50%.

Table 1: List of differences of alginate lyase.

No. Source Gene PL

Molar

mass

(kDa)

Alginate

lyase

specificity

Cleavage

mode

Exo/Endo

Temperature

(℃)

Activity

(U/mg)

Reference

Nitratiruptor sp.

SB155-2

NitAly 7 29 pM Endo 70 6 1620

(Inoue,

2016)

Streptomyces

coelicolo

A3(2)

OUC-

ScCD6

6 52 pM Endo 50 9 31.6

(Cheng,

2020)

3 Vibrio sp. W13 Algb 7 55.05 Alg, pMG Endo 30 8 457

(Zhu,

2015b)

Haliotis discus

hanna

HdAly 14 28.9 pM Endo 45 8 28.9

(Shimizu,

2003)

Flavobacterium

sp. S02

AlyS02 7 36.5 pM, pG Endo 30 7.6 442.84

(Zhou,

2020)

Flavobacterium

sp. H63

rSAGL 7 32 pM Endo 45 7.5 4044

(Li,

2018)

Pseudomonas sp.

E03

AlgA 5 40.4 pM Endo 30 8 222

(Zhu,

2015a)

8 Vibrio sp. SY01 Aly08 7 35 pG Endo 45 8.35 841

(Wang,

2019)

Zobellia

alactanivorans

AlyA5 7 42 pG Exo - 7 449.3

(Thomas,

2013)

carrageenovora

ASY5

Aly1281 7 40.65 pM, pG Endo 50 8 1.15

(Zhang,

2020)

Shewanella sp.

YH1

rAlgSV1-

PL7

7 33.22

Alg, pM,

Endo+

Exo

45 8 96

(Yagi,

2017)

Haliotis discus

hanna

HdAlex 14 32 pM Exo 42 7.1 684

(Suzuki,

2006)

3 APPLICATION

Degradation of alginate into oligosaccharides is the

current major trend and key step in the use of

seaweed polysaccharides. Enzymatic hydrolysis

prepared by different alginate lyases, the products

are also different. As early as the 1990s, studies have

shown that alginate oligosaccharides could promote

growth of plant roots. Yoshihiba et al. prepared

alginate oligosaccharides by enzymatic hydrolysis.

According to the experimental results, it was

confirmed that alginate oligosaccharides had a

significant growth-promoting effect on plant

rhizomes. In 2020, a study conducted by

Aitouguinane et al. pointed out that alginate

polysaccharides and its oligosaccharides derivatives

ICBEB 2022 - The International Conference on Biomedical Engineering and Bioinformatics

trigger plant defense responses and show a

significant ability to induce PAL activity in tomato

seedling leaves(Aitouguinane, et al., 2020). These

results help marine life as a potential biological

resource to protect plants from plant pathogens in the

context of ecologically sustainable green technology.

Studies have found that extracellular polysaccharides

are important pathogenic factors for some

pathogenic bacteria to cause lung infections. It is

well known that alginate exists in biofilms as a key

role in the failure of immunotherapy and antibiotic

therapy. In 2020, a study by wan et al. reported a new

silver nanocomposite, which is used to deliver drug

compounds, alginate lyase and ceftazidime. Due to

the principle of alginate degradation of alginate

lyase, it has a strong inhibition and degradation effect

on the Pseudomonas aeruginosa PAO1, and it can

also eradicate Pseudomonas aeruginosa in the lung

to achieve the purpose of treatment(Wan, 2020).

With the consumption of petroleum and other

non-renewable energy, the development of new

clean energy has been put on the agenda. There have

been many articles reporting from alginate. The main

principle of the research progress in the preparation

of bioethanol from alginate is to prepare bioethanol

through the synergistic action of the alginate lyase

endonuclease and exonuclease and continuous

reaction saccharification of alginate.

4 CONCLUSION

Alginate lyases have become new tool enzymes

because of its unique biological activity and high

efficiency, clean and controllable properties.

Recently, the research on alginate lyases have

gradually shifted from the early description of its

characteristics to the biological study of its

mechanism. Meanwhile, with the rapid development

of molecular biology, a variety of alginate lyase

genes can be cloned, sequenced and selected for

heterologous expression to construct efficient

engineering strains. It can be predicted that the

application of alginate lyases will be more and more

extensive in the future.

ACKNOWLEDGMENTS

This work was supported by the Key Technology

Research and Development Program of Shandong

Province [grant numbers 2019QYTPY024,

2019YYSP019]; Key Technology Research and

Development Program of China [grant number 2020-

CXY45]; Spring Industry Leader Talent Support

Plan [grant numbers 2017035, 2019042]; Shandong

Taishan Leading Talent Project [grant numbers

LJNY202015, tscy20180507]; Shandong Provincial

Natural Science Foundation (No. ZR2020QC008);

and Science, Education, and Industry Integration

Innovation pilot project at Qilu University of

Technology (Shandong Academy of Sciences) [grant

numbers 2020KJC-YJ01, 2020KJC-GH10].

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