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Biotechnological Production of the Sunscreen Pigment Scytonemin in Cyanobacteria: Progress and Strategy

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Biotechnological Production of the Sunscreen Pigment Scytonemin in Cyanobacteria: Progress and Strategy marine drugs Review Biotechnological Production of the Sunscreen Pigment Scytonemin in Cyanobacteria: Progress and Strategy Xiang Gao 1,* , Xin Jing 1, Xufeng Liu 2 and Peter Lindblad 2,* 1 School of Food and Biological Engineering, Shaanxi University of Science & Technology, Xi’an 710021, China; [email protected] 2 Microbial Chemistry, Department of Chemistry-Ångstrom, Uppsala University, Box 523, 751 20 Uppsala, Sweden; [email protected] * Correspondence: [email protected] (X.G.); [email protected] (P.L.) Abstract: Scytonemin is a promising UV-screen and antioxidant small molecule with commercial value in cosmetics and medicine. It is solely biosynthesized in some cyanobacteria. Recently, its biosynthesis mechanism has been elucidated in the model cyanobacterium Nostoc punctiforme PCC 73102. The direct precursors for scytonemin biosynthesis are tryptophan and p-hydroxyphenylpyruvate, which are generated through the shikimate and aromatic amino acid biosynthesis pathway. More upstream substrates are the central carbon metabolism intermediates phosphoenolpyruvate and erythrose- 4-phosphate. Thus, it is a long route to synthesize scytonemin from the fixed atmospheric CO2 in cyanobacteria. Metabolic engineering has risen as an important biotechnological means for achieving sustainable high-efficiency and high-yield target metabolites. In this review, we summarized the biochemical properties of this molecule, its biosynthetic gene clusters and transcriptional regulations, the associated carbon flux-driving progresses, and the host selection and biosynthetic strategies, with the aim to expand our understanding on engineering suitable cyanobacteria for cost-effective Citation: Gao, X.; Jing, X.; Liu, X.; production of scytonemin in future practices. Lindblad, P. Biotechnological Production of the Sunscreen Pigment Keywords: cyanobacteria; sunscreen pigments; scytonemin; metabolic engineering Scytonemin in Cyanobacteria: Progress and Strategy. Mar. Drugs 2021, 19, 129. https://doi.org/ 10.3390/md19030129 1. Introduction Cyanobacteria are photoautotrophic prokaryotes that can directly convert CO2 into Academic Editor: Chingfeng Weng organic compounds using solar energy. Cyanobacteria possess relatively small genomes and can grow with minimal nutrient requirement. Engineering cyanobacteria offers an Received: 4 February 2021 attractive approach to drive carbon flux to the biosynthesis of fuels, chemicals, medicines, Accepted: 24 February 2021 Published: 27 February 2021 plant secondary metabolites, and other value-added products [1–5]. Cyanobacteria naturally grow in diverse habitats including fresh water, oceans, al- Publisher’s Note: MDPI stays neutral kaline waters, cold deserts, polar regions and hot springs. To cope with the extreme and with regard to jurisdictional claims in changeable environments, cyanobacteria have developed a wide range of adaptive mecha- published maps and institutional affil- nisms. Some cyanobacteria are directly exposed to strong sunlight in native habitats, such iations. as those dwelling in intertidal mats or on desert soil surfaces [6–8]. Intense ultraviolet (UV) radiation can cause cell damage by both direct effects on proteins and nucleic acids and indirect effects via the induced reactive oxygen species [9]. To eliminate the deleterious effects of UV radiation, one of the most important strategies in some cyanobacteria is to generate UV-absorbing compounds, such as mycosporine-like amino acids (MAAs) Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. and scytonemin [10]. Both compounds are located in the exopolysaccharide matrix and This article is an open access article constitute the first line of defense against UV penetration and subsequent damage [11–13]. distributed under the terms and Multiple estimates from the fossil record of cyanobacteria and relaxed molecular clock conditions of the Creative Commons models suggest a minimum age for the evolutionary advent of scytonemin at around Attribution (CC BY) license (https:// 2.1 billion years [14]. creativecommons.org/licenses/by/ Scytonemin is naturally biosynthesized in some cyanobacteria and has a great prospect 4.0/). in cosmetic and biomedical application. However, as far as we know, enhanced production Mar. Drugs 2021, 19, 129. https://doi.org/10.3390/md19030129 https://www.mdpi.com/journal/marinedrugs Mar. Drugs 2021, 19, x 2 of 18 Mar. Drugs 2021, 19, 129 2 of 16 Scytonemin is naturally biosynthesized in some cyanobacteria and has a great pro- spect in cosmetic and biomedical application. However, as far as we know, enhanced pro- duction of this compound in cyanobacteria via metabolic engineering has not been re- of this compound in cyanobacteria via metabolic engineering has not been reported. To ported. To exploit the maximal potential of engineering cyanobacterial hosts for exploit the maximal potential of engineering cyanobacterial hosts for scytonemin biosyn- scytonemin biosynthesis is the next important step. In this review, we summarized the thesis is the next important step. In this review, we summarized the research progresses research progresses regarding scytonemin, placing emphasis on its biosynthetic route and regarding scytonemin, placing emphasis on its biosynthetic route and the metabolic engi- the metabolic engineering technologies from cyanobacteria and other microorganisms, neering technologies from cyanobacteria and other microorganisms, with the aim to expand with the aim to expand our understanding on metabolically engineering of cyanobacterial our understanding on metabolically engineering of cyanobacterial hosts for cost-effective hosts for cost-effective production of scytonemin in future practices. production of scytonemin in future practices. 2. 2.Functional Functional Roles Roles of ofScytonemin Scytonemin 2.1.2.1. Molecular Molecular Structure Structure and and UV UV Absorption Absorption ScytoneminScytonemin is isa lipid a lipid soluble soluble and and yellow-brown yellow-brown pigment pigment exclusively exclusively found found in insome some cyanobacteriacyanobacteria [11]. [11 ].This This compound compound is isa dimer a dimer composed composed of ofindolic indolic and and phenolic phenolic subunits subunits symmetricallysymmetrically connected connected through through a carbon–car a carbon–carbonbon bond bond (Figure (Figure 1).1 ).Scytonemin Scytonemin has has oxi- oxi- dizeddized (yellow; (yellow; Mw Mw 544 544 Da) Da) and and reduced reduced (red; MwMw 546546 Da)Da) forms forms and and also also three three derivatives, deriva- tives,dimethoxyscytonemin, dimethoxyscytonemin, tetramethoxyscytonemin, tetramethoxyscytonemin, and and scytonin scytonin [15, 16[15,16].]. FigureFigure 1. The 1. The scytonemin scytonemin molecule molecule (A) and (A) andits 3D its model 3D model (B). Two (B). images Two images are generated are generated by the by the MolViewMolView program program (https://molview.o (https://molview.org/rg/, accessed, accessed on 20-Aug-2020). on 20 August 2020). TheThe solar solar UV UV radiation radiation can can be be divided divided into into UV-A UV-A (315–400 (315–400 nm), nm), UV-B UV-B (280–315 (280–315 nm), nm), andand UV-C UV-C (100–280 (100–280 nm). nm). UV-C UV-C does does not not reach reach the the earth’s earth’s surface surface owing owing to to stratospheric stratospheric ozone.ozone. The The remaining remaining UV UV that that reaches reaches the the surf surfaceace of ofthe the earth earth consists consists of of95% 95% UV-A UV-A and and 5%5% UV-B UV-B [17]. [17 ].Scytonemin Scytonemin can can absorb absorb the the th threeree UV UV radiations, radiations, but but mainly mainly the the UV-A, UV-A, and and cancan prevent prevent about about 90% 90% of UV-A of UV-A from from entering entering the cells the cells[11,18,19]. [11,18 It,19 can]. Italso can provide also provide sub- stantialsubstantial protection protection against against UV-B and UV-B UV-C and damage UV-C damage [20,21]. [ 20The,21 absorption]. The absorption in the UV-C in the rangeUV-C implies range that implies scytonemin that scytonemin is an ancien is ant pigment ancient possibly pigment evolved possibly during evolved the during Precam- the brianPrecambrian [20]. Scytonemin [20]. Scytonemin has an in hasvivo an maximumin vivo maximum absorption absorption at 370 nm atand 370 an nm in andvitro an maximumin vitro maximum absorption absorption at 384 nm at[11,22]. 384 nm [11,22]. 2.2. Solubility and Stability 2.2. Solubility and Stability Scytonemin does not dissolve in water, but dissolves readily in lipids. In liquid sus- Scytonemin does not dissolve in water, but dissolves readily in lipids. In liquid sus- pension culture of Nostoc flagelliforme, yellow-brown scytonemin was observed to be tightly pension culture of Nostoc flagelliforme, yellow-brown scytonemin was observed to be associated with the released exopolysaccharides [23]. Scytonemin was usually extracted from tightly associated with the released exopolysaccharides [23]. Scytonemin was usually ex- samples with solvents such as 100% acetone or 100% ethyl acetate [11], 100% acetonitrile [24], tracted from samples with solvents such as 100% acetone or 100% ethyl acetate [11], 100% 80% tetrahydrofuran [13], and methanol:ethyl acetate (1:1, v/v)[9,21]. In addition, we found acetonitrile [24], 80% tetrahydrofuran [13], and methanol:ethyl acetate (1:1, v/v) [9,21]. In scytonemin was soluble in N,N-dimethylformamide and the polyvinylpyrrolidone K30 addition,aqueous we solution. found scytonemin Its solubility
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