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. 2025 Aug 7;15:28882. doi: 10.1038/s41598-025-14788-1

Evaluation of how laser photostimulation at two wavelengths alters the antimicrobial potential of Streptomycetes spp

Noor Abduljabar Jadah 1,✉, Talib Saleh Al-Rubaye 2
PMCID: PMC12332045  PMID: 40775007

Abstract

The purpose of this study was to isolate and identify additional products produced by direct laser irradiation, as well as to ascertain if laser irradiation may stimulate the synthesis of antibiotic compounds in a local Streptomycetes (Strept). Moreover, we postulate the mechanisms by which lasers function within living bacterial cells and suggest that sequential photochemical reactions may transpire following a designated period of irradiation. Thiophene was found as one of the most significant clinical products with antibacterial and anticancer properties. To accomplish these objectives, we selected two isolates: Streptomyces thinghirensis strain S10 (Strept thin), which inherently synthesizes an antibacterial agent, and Streptomyces lienomycini strain C.P.57 (Strept.lieno), which does not generate antimicrobials. The experimental isolates were exposed to identical circumstances as the control isolates, with the exception that the inoculum underwent irradiation with a diode laser for varying durations. We initially assessed the antibacterial efficacy of the irradiation and control Strept. Gas chromatography-mass spectrometry (GC-MS) was utilized to discover antibacterial substances. Ultimately, we determined that laser irradiation caused alterations in both antimicrobial-producing and non-producing Strept.; specifically, those that produced antimicrobials ceased to do so post-irradiation, and conversely.

Supplementary Information

The online version contains supplementary material available at 10.1038/s41598-025-14788-1.

Keywords: Streptomycetes, Laser, Thiophene, Gas-chromatography, Antibacterial, Antitumor

Subject terms: Industrial microbiology, Biotechnology, Drug discovery, Medical research

Introduction

Streptomyces are Gram-positive filamentous bacteria that are found naturally in organic soil. These bacteria are among the most important bacteria in clinical drug manufacturing, as they are considered a major source of a vast range of antibiotics, and anticancer reagents1. The ability of bacteria to produce certain types of products depends on many factors, such as the environment of the isolate, growth conditions, species or genus and the physiological and metabolic status2. Therefore, many studies have focused on identifying the optimal conditions to induce the production of desired products in Strept. For example, in a study by Al-Rubaye et al. in 2020, different types of antibiotics and bioactive compounds produced by several Strept isolates obtained from the Tigris River in Baghdad, were identified by gass chromatoghrphy(GC)3,4. Many studies have focused on modulating the secondary metabolites of Strept by using fermentation media with limited organic nitrogen contents5–7. Furthermore, many scientific publications confirm that the types of secondary metabolites present depend mainly on the particular species of Strept owing to the different cell wall structure, and certain genetic modifications found within this genus, which is the largest recognized taxonomic item in the phylum Actinomycetes8.

Among the most significant bioactives compounds produced by certain of Strept are thiophenes, which are heterocyclic metabolites commonly produced by both plant species belonging to the Asteraceae family and filamentous Strept bacteria9. These metabolites have considerable bioactivities, such as antimicrobial, antiviral, anti-inflammatory, antioxidant, insecticidal, and antitumor properties9,10. The chemical name of thiophene is thiacyclopentadiene, with a formula of C4H4S, where the sulfur atom in the aromatic rings has a significant role in medical chemistry, and can form bonds with different atoms, such as carbon, nitrogen, oxygen, phosphorus and halogens. Therefore, sulfur-based functionalities have become important pharmacophores in drug discovery for the design and synthesis of new derivatives and analogues11.

Numerous experiments have investigated, the production of thiophene derivatives from Strept via precursor-based combinatorial biosynthesis. Recently, a thiophene precursor was added to Strept fermentation broth, and the product was identified via nuclear magnetic resonance and mass spectrometry12. In this study, we aimed to determine if laser irradiation could stimulate the biosynthesis of thiophene in a local Strept isolate.

Methods

Microorganisms

The Strept isolates were obtained from the Department of Biotechnology/ College of Science at the University of Baghdad13. The Isolates were selected, purified and identified biochemically. Both the Streptomyces lienomycini strain C.P.57 (Strept. lieno) and Streptomyces thinghirensis strain S10 (Strept thin) were identified at the species level using 16 S rRNA gene sequencing.

In this study we used S. aureus and E. coli as model Gram-positive and gram-negative bacteria, respectively, which were subjected to antibacterial sensitivity tests with the Strept crude extracts.

Media and broth

In this study we used casein starch media as a solid medium for slant storage and Strept cultivation prior to colony counting. Casein starch broth, which consisted of 10gm of starch, 3 g of casein, 0.02 g of CaCO3, 0.01 g of Fe3SO4.7H2O, 2 g of KNO3, 0.05 g of MgSO4.7H2O, 2 g of NaCl in 100 ml distilled water, was used in the antimicrobial activity assays. Sterile broth inoculated with a full loop of Strept was inoculated in 20 ml of production broth at pH 7.2 in a 100 ml conical flask, and incubated at 30 °C and with shaking at 150 rpm for 10 days 13–15.

Soybean casein digest broth (HIMEDIA) was used to activate the pathogenic bacteria via 18–24 h of incubation at 37 °C.

Irradiation procedure

In this experiment, we used two devices equipped with diode lasers (JD-R303, HUONJE 114 TM/ China) one laser emitted at a wavelength of 650 nm and the other at a wavelength of 532 nm. The other parameters of the two devices were the same, as shown in Table 1 :

Table 1.

Irradiation device specifications.

The parameters Dose
Laser power 100mW
Type of proliferation CW
Time of exposure 1 min, 2 min, 3 min
Diameter of laser beam 0.7 mm
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The laser apparatus setup and irradiation procedure were described previously (Jadah 2022)16.A fresh culture of Strept was incubated for 10 days at 30 °C in a rotating incubator; under sterile conditions. Each sample was diluted to 103 cells/ml before irradiation, in the irradiated and nonirradiated (control) groups to ensure that all the cells and spores received an equal dose of laser irradiation.

The samples were divided into 7 groups, with 5 replicates in each group, as follows:

  1. Non-irradiated group(control) (5 flasks).

  2. Three groups of Strept irradiated at 650 nm three times (15 flasks).

  3. Three groups of Strept at 532 nm three times (15 flasks).

Laser irradiation of 1 ml of cell suspension (103 cells/ml), was performed in a dark room at a power of 100 mW at 650 nm for 1 min (for example), and then immediately inoculated into a 100 ml flask containing 20 ml of production broth. The inoculated flasks were transferred immediately to a shaker incubator, for incubation at temperature 30 °C and 150 rpm for 10 days.

Crude extraction

After 10 days of shaking incubation, 10 ml of liquid bacterial culture was transferred to test tubes and centrifuged at 8000 rpm for 10 min. Then, a portion of the supernatant was used for antimicrobial sensitivity testing via the well diffusion method. Another 10 ml of the supernatant was lyophilized with a bench-top freeze dryer (LYO60B-1PT, Infitek Company) to produce a solid powder for GC-MS analysis.

Antibacterial sensitivity test

Mueller‒Hinton agar plates were divided into two groups; the S. aureus group and the E. coli group. Each plate was inoculated with 0.1 ml of pathogenic bacteria (103 cells/ml) by spreading with sterile cotton swabs for 18 h of incubation. Then, wells ( 6 mm in diameter) were made in each plate with sterile micropipette tips. The crude supernatant (0.1 ml) was added to each well, and the plates were incubated for 24 h at 37 °C.

Statistical analysis

The results indicate the diameters of radius (in cm) of the inhibitory growth zones surrounding the wells in the pathogenic bacterial cultures (E. coli and S. aureus). The collected data were analyzed using GraphPad Prism version 10.5.0. The comparisons were conducted between the control and irradiated groups using standard One-Way ANOVA to demonstrate the significant differences among means (p < 0.05) of the maximum values of antibacterial potentials in the irradiation groups. The maximal antibacterial potentials were assessed by T-test analysis by comparing the means of irradiated and non-irradiated groups for each Strept strain.

GC-MS analysis

The supernatants were examined utilizing GC–MS. (7820 A GC system, Agilent Technologies) (Table 2). The mass spectra of the lyophilized crude materials were compared with spectra of known compounds in the National Institute Standard and Technology (NIST) database to determine the name, molecular formula and weight, and the area under the curve of the compounds17. The specification of GC-Mass system that were used in identifying produced compounds were listed in Table 2.

Table 2.

Instrument conditions of GC-MS analysis.

No. Parameter descriptions
1 Gas Chromatograph Agelint Technologies(7820 A)
2 GC Mass Spectrometer (5977E) USA
3 Analytical Column Agelint HP-5ms Ultra lneit (30 m length x 250 μm inner diameter x 0.25 μm film thickness)
4 Injection volume 1 µl
5 Pressure 11.933 psi
6 GC Inlet Line Temperature 250 ˚C
7 Aux heateres Temperature 300 ˚C
8 Carrier Gas He 99.99%
9 Injector Temperature 250 ˚C
10 Scan Range m/z 25-1000
11 Injection Type Splitless
12 Oven Program

Temperature

Ramp 1 60 ˚C hold to 3 min.

Ramp 2 60 ˚C to 180 ˚C 7 ˚C/min

Ramp 3 180˚C to 280˚C 8 ˚C/min

Ramp 4 280˚C hold to 5 min.
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Results

Antibacterial potential of the isolates

The Strept. lieno isolates exhibited smaller inhibition zones with the test pathogenic bacteria before laser irradiation. These isolates exhibited a lighter pink color compared to their non-irradiated counterparts in the casein starch medium following laser irradiation, whereas the Strept thin isolates lost their pigmentation after irradiation. First, the antimicrobial potential of the Strept. isolates against S. aureus and E. coli was examined via the well diffusion method and measurement the radius of the inhibition zone. These data were analyzed by GraphPad Prism, as shown in Fig. 1.

Fig. 1.

Fig. 1

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Comparisons of the mean values between non- irradiated and irradiated Strept. groups; where the non-irradiated groups (control) include; Strept thin: Streptomyces thinghirensis strain S10, and Strept. lieno: Streptomyces lienomycini strain C.P.57. Strept thin E coli 532/3: antibacterial potential of the irradiated Strept thin with 532 nm laser wavelength for 3 min of exposure time against E coli. Strept thin S. aureus 532/2: antibacterial potential of the irradiated Strept thin with 532 nm laser wavelength for 2 min of exposure time against S.aureus; Strept. lieno E coli650/2: antibacterial potential of the irradiated Strep. lieno with 650 nm laser wavelength for 2 min of exposure time against E coli; Strept. lieno S. aureus 532/2: antibacterial potential of the irradiated Strept lieno with 532 nm laser wavelength for 2 min of exposure time against S.aureus.

Figure 1 indicates that the antibacterial efficacy of the non-irradiated group, Strept thin, exceeds that of Strept lieno, albeit not significantly. The effect of laser irradiation on the antibacterial properties of Strept thin occurred at a wavelength of 532 nm, whereas Strept lieno responded to both wavelengths at the same exposure duration. The antibacterial impact of irradiated Strept. lieno on E. coli and S. aureus is significantly greater than that of irradiated Strept. thin. The inhibitory effect of Strept. lieno following 532 nm irradiation was effective against both bacterial strains. The peak inhibitory impact against E. coli was almost 95% at a wavelength of 650 nm, whereas the efficacy against S. aureus was approximately 86% following irradiation at 532 nm.

GC–MS

To identify the antibacterial compounds produced by the Strept strains before and after laser irradiation that inhibited the growth of S. aureus and E. coli, we utilized GC‒MS. Considering the NIST library, the GC–MS peaks are shown in Figs. 2 and 3. The area percentages of the peaks allowed semiquantitative analysis of metabolite abundance, as shown in Tables 3 and 4.

Fig. 2.

Fig. 2

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Curve of secondary metabolites produced by Strept. lieno (control) as it obtained from software of GC-mass spectroscopy machine. The y-axes represent the abundance of magnetic field that will change relatively according to different biomolecules in lyophilized extracellular crude of Strept. Lieno, that pass through heated thin coil-canals. The x-axes is the time (-second) for biomolecules respond to magnetic field.

Fig. 3.

Fig. 3

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Curve of secondary metabolites produced by Strept. lieno (irradiated with 650 nm for 2 min) as it obtained from software of GC-mass spectroscopy machine.

Table 3.

The molecular components of extracellular crude powder of non-irradiated Strept. lieno18–26.

Peak no Retention time Area (%) Library (ID) Reference peak CAS number Qualitative (magnetic susceptibility) Description and importance
1 5.134 4.64 Ethyl formate 806 000109-94-4 43 Secondary metabolite that is an ester formed when ethanol reacts with formic acid (18)
Ethylene oxide 73 000075-21-8 12 Secondary metabolite isomer of acetaldehyde and vinyl alcohol18
Ethylene oxide 75 000075-21-8 16 Secondary metabolite isomer of acetaldehyde and vinyl alcohol18
2 6.398 1.42 (+/−)-2-Amino-1-propanol 905 006168-72-5 12 Contains both amine (− NH2) and alcohol (− OH) groups and exhibits characteristics of both19,20
(+/−)-2-Amino-1-propanol 904 006168-72-5 12 Contains both amine (− NH2) and alcohol (− OH) groups and exhibits characteristics of both19,20
Cyclobutanol 663 002919-23-5 14 Hydrogen bond donor and starting material for the production of pharmaceuticals, including antimicrobial agents and drugs for treating cancer19
3 6.736 3.51 4-Methyl-1,3-dioxolane 2052 001072-47-5 27 No application was reported
Octanoic acid methyl ester 30,223 000111-11-5 25 Extracellular membrane fatty acid18,21
Octanoic acid methyl ester 30,233 000111-11-5 14 Extracellular membrane fatty acid18,21
4 8.311 1.52 N-Methylglycine 2163 000107-97-1 16 Amino acid also known as sarcosine18
Propanamide 728 000079-05-0 14 No application was reported
Propanamide 726 000079-05-0 14
5 11.470 1.51 (3-Hydroxy-5-methyl-2-oxo-2,3-dihydro-1 H-indole-3-carbonyl) urea 101,141 1000296-21-6 9 No application was reported
Hexamethyl disilathiane 44,259 003385-94-2 14 Flammable, highly toxic irritant18,22
3-(2 Benzoxazolylthio)-1-phenyl-propenone 128,273 299461-72-6 14 Cyclooxygenase-2 (COX-2) inhibitory activity19,23
6 12.084 1.13 Cyclobutanol 655 002919-23-5 58 Hydrogen bond donor and starting material for the production of pharmaceuticals, including antimicrobial agents and drugs for treating cancer19
(+/−)-2-Amino-1-propanol 906 006168-72-5 42 Contains both amine (− NH2) and alcohol (− OH) groups and exhibits characteristics of both10,20
Cyclobutanol 663 002919-23-5 52 Hydrogen bond donor and starting material for the production of pharmaceuticals, including antimicrobial agents and drugs for treating cancer19
7 14.369 1.56 N-[3,5-Dinitropyridin-2-yl] proline 128,542 003264-09-3 4 No application was reported
Sarcosine, N- valeryl pentadecyl ester 204,024 1000321-56-8 4 No application was reported
Sarcosine, N-valeryl pentadecyl ester 204,024 1000321-56-8 4 No application was reported
8 20.385 22.44 Hexadecanoic acid methyl ester 119,400 000112-39-0 96 Fatty acid methyl ester18–24
Hexadecanoic acid methyl ester 119,407 000112-39-0 95 Fatty acid methyl ester24
Hexadecanoic acid methyl ester 119,408 000112-39-0 91 Fatty acid methyl ester18,24
9 20.385 22.44 Propanamide 728 000079-05-0 47 No application was reported
N,2-Dimethyl-1-propanamine 1935 000625-43-4 46 No application was reported
4-Chloro-alpha methyl-benzeneethanamine 37,816 000064-12-0 38 Irritant and environmental hazard, secondary metabolite18,25
10 21.233 0.9 Chloroamphetamine 37,813 1000248-75-1 38 Replacement monoamine and amphetamine releaser neurotoxic to serotonergic neurons only20,21
4-Chloro-alpha methyl-benzeneethanamine- 37,816 000064-12-0 47 No application was reported
DL-Phenylephrine 36,227 001477-63-0 49 Alpha-1 adrenergic agonist and irritating substance that is typically given in conjunction with anesthesia during surgery to treat hypotension26
11 22.505 4.6 (Z, Z)-9,12-Octadecadienoic acid 127,648 000060-33-3 91 Doubly unsaturated fatty acid18
Trans-10-Methyl-12-cis octadecadienoate 139,709 1000336-44-2 94 Fatty acid18
9,12-Octadecadienoic acid methyl ester 139,708 002462-85-3 96 Fatty acid18
12 22.601 30.05 (Z)-9-Octadecenoic acid methyl ester 141,302 000112-62-9 93 Fatty acid18
11-Octadecenoic acid methyl ester 141,290 052380-33-3 91 Fatty acid18
9-Octadecenoic acid methyl ester, (E)- 141,310 001937-62-8 99 Fatty acid18
13 22.912 7.69 Methyl stearate 143,126 000112-61-8 99 Fatty acid18
Methyl stearate 143,128 000112-61-8 95 Fatty acid18
Methyl stearate 143,130 000112-61-8 83 Fatty acid18
14 23.155 6.1 Metaraminol 36,216 000054-49-9 38 Secondary metabolite sympathomimetic that is used to prevent and treat hypotension, which is a common side effect of anesthesia26
Racepinephrine 48,249 000329-65-7 38 Short-term remedy for moderate asthma symptoms18
6-Methyl-2-piperidinone 6942 004775-98-8 41 Can be utilized to synthesize vitamin K1, vitamin A, vitamin E, and a variety of molecules with various tastes and scents25
15 23.683 1.3 Metaraminol 36,216 000054-49-9 46 Secondary metabolite sympathomimetic that is used to prevent and treat hypotension, which is a common side effect of anesthesia26
Meglumine 57,369 006284-40-8 43 Glucose-derived sugar alcohol18
Meglumine 57,368 006284-40-8 43 Glucose-derived sugar alcohol18
16 25.180 2.42 Methyl 18-methylnonadecanoate 166,215 1000352-20-6 99 Fatty acid18
Eicosanoic acid methyl ester 166,219 001120-28-1 92 Fatty acid18
Eicosanoic acid methyl ester 166,218 001120-28-1 90 Fatty acid18
17 26.279 0.85 Metaraminol 36,216 000054-49-9 49 Secondary metabolite sympathomimetic that is used to prevent and treat hypotension, which is a common side effect of anesthesia26
L-Alanine-4-nitroanilide 68,810 001668-13-9 49 Secondary metabolite used as a substrate for alanine aminopeptidase (AAP) activity determination24
Phenylephrine 36,221 000059-42-7 49 Alpha-1 adrenergic agonist and irritant that is typically given in conjunction with anesthesia during surgery to treat hypotension26
18 27.257 1.07 N-Methyl-1-octanamine 20,375 002439-54-5 38 Fatty acid18
Cotinine 23,581 071031-15-7 25 Monoamine alkaloid monoamine alkaloid found in the shrub Catha edulis (khat)26
Cyclobutanol 655 002919-23-5 38 Hydrogen bond donor and starting material for the production of pharmaceuticals, including antimicrobial agents and drugs for treating cancer19
19 29.118 2.62 Trimethyl[4-(2-methyl-4-oxo-2-pentyl)phenoxy]silane 113,997 1000283-54-9 16 Anti-inflammatory, antioxidant and antibacterial Compound19
2,4,6-Cycloheptatrien-1-one, 3,5-bis-trimethylsilyl- 102,104 1000161-21-8 22 Antioxidant, anticancer19
Trimethyl[4-(1-methyl-1-methoxyethyl)phenoxy]silane 92,131 1000283-54-8 12 Anti-inflammatory, antioxidant and antibacterial compound19
20 33.740 0.9 Ala-.beta.-Ala, trimethylsilyl ester 87,953 1000333-69-0 27 No application was reported
Fluoxetine 151,963 054910-89-3 22 Utilized to treat panic disorder, bulimia nervosa, depression, premenstrual dysphoric disorder (PMDD), and obsessive-compulsive disorder (OCD)26
1-Octadecanamine, N-methyl- 130,249 002439-55-6 30 No application was reported
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Table 4.

The molecular components of extracellular crude powder of irradiated Strept. lieno18–26.

Peak no. Ratio Area% Library/ID Beilsten refrence CAS number Magnetic susceptibility Description and importance
1 5.160 6.91 Hydrazinecarboxamide 908 000057-56-7 9 Known as Aminourea it is irritate and acute toxic compound (toxin) 18.
Ethylene oxide 75 000075-21-8 12 Secondary metabolites that is isomeric with acetaldehyde and vinyl alcohol18.
Cyacetacide 3437 000140-87-4 9 Known as Cyanoacetohydrazide it is irritate and acute toxic compound(toxin) 18.
2 5.454 3.30 1-Propanol, 2-amino-, (.+/-.)- 663 002919-23-5 10 The compound possesses both the amine group -NH2 and the alcohol group -OH, thus exhibiting characteristics of both groups18.
1-Propanol, 2-amino-, (.+/-.)- 289 000071-23-8 10
1-Propanol, 2-amino-, (.+/-.)- 90 006168-72-5 10
3 6.173 2.96 Cyclobutanol 663 002919-23-5 10 Cyclobutanol serves as a hydrogen-bond donor and is used as a starting material for the production of pharmaceuticals, including antimicrobial agents and drugs for treating cancer 19.
1-Propanol 289 000071-23-8 10 Alcohol used as a solvent and chemical intermediate18.
Cyclobutanol 655 002919-23-5 11 Cyclobutanol serves as a hydrogen-bond donor and is used as a starting material for the production of pharmaceuticals, including antimicrobial agents and drugs for treating cancer 19.
4 6.744 6.45 Propanamide, N-(1cyclohexylethyl) 47,937 1000142-14-3 38 No application was reported
Guanidine, N,N-dimethyl- 1860 006145-42-2 18 No application was reported
Cyclobutanol 655 002919-23-5 11 Cyclobutanol serves as a hydrogen-bond donor and is used as a starting material for the production of pharmaceuticals, including antimicrobial agents and drugs for treating cancer 19.
5 7.921 1.61 Cyclobutanol 655 002919-23-5 46
Cyclobutanol 663 002919-23-5 43
2-Hexanamine 4135 005329-79-3 47 No application was reported
6 8.198 1.72 Cyclotetrasiloxane, octamethyl- 141,481 000556-67-2 38 Organo-silicon substance cause hazard for human and environment (toxin) 18.
Cyclotetrasiloxane, octamethyl- 141,484 000556-67-2 78
1,1,3,3,5,5,7,7-Octamethyl-7-(2-methylpropoxy)tetrasiloxan-1-ol 196,332 1000364-61-2 35 Organo-silicon compound 18.
7 8.302 1.46 Cyclobutanol 663 002919-23-5 14 Cyclobutanol serves as a hydrogen-bond donor and is used as a starting material for the production of pharmaceuticals, including antimicrobial agents and drugs for treating cancer19.
2-Aminononadecane 130,243 031604-55-4 10 No application was reported
Cyclobutanol 655 002919-23-5 11 Cyclobutanol serves as a hydrogen-bond donor and is used as a starting material for the production of pharmaceuticals, including antimicrobial agents and drugs for treating cancer19.
8 11.496 3.18 Thiophene, 2-(bromoacetyl)-5-[bis(dimethylamino)phosphinoyl]- 174,743 1000142-86-5 33 antiviral, antioxidant, antibacterial, anti-inflammatory, insecticidal, and anti-tumor 10,19.
Acetamide, 2-cyano- 1335 000107-91-5 4 The precursor reagent for the synthesis of vitamin B6 is cyanoacetamide20.
2-(4,5-Dihydro-3-methyl-5-oxo-1-ph enyl-4-pyrazolyl)-5-nitrobenzoic acid 194,638 020307-76-0 4 Acute toxin18.
9 14.378 1.80 N-[3,5-Dinitropyridin-2-yl]proline 2,128,542 003264-09-3 2 Bioactive compound 21.
Sarcosine, N-valeryl-, pentadecyl ester 204,024 1000321-56-8 4 Amino acid derivatives18.
Sarcosine, n-hexanoyl-, pentadecyl ester 210,813 1000321-13-0 2 Amino acid derivatives18.
10 16.974 1.38 4-Pyridinecarboxamide, 6-chloro-4, 5-dicyano-2-[(cyclohexylidenamino) oxy]-1,2,3,4-tetrahydro-3,3-dimeth yl- 183,188 1000350-38-4 2 derivative of isonicotinic acid 18.
N-[3,5-Dinitropyridin-2-yl]proline 128,542 003264-09-3 1 Bioactive compound 21.
3,6-Bis-dimethylaminomethyl-2,7-di hydroxy-fluoren-9-one 165,814 1000318-33-0 12 No application was reported
11 19.485 1.42 Cyclobutanol 663 002919-23-5 53 Cyclobutanol serves as a hydrogen-bond donor and is used as a starting material for the production of pharmaceuticals, including antimicrobial agents and drugs for treating cancer 19.
2-Butanamine, 3-methyl- 1914 000598-74-3 50 Acute toxic18.
2-Ethoxyamphetamine 44,804 135014-84-5 64 Biomolecule similar to a drug of the amphetamine class22.
12 20.393 19.53 Hexadecanoic acid, methyl ester 119,400 000112-39-0 99 Type of fatty acids, used as Hepatoprotective; hypocholesterolemic; antifungal; antiarthritic; antitumor; anticancer; anticoronary; anti-inflammatory compound23.
Hexadecanoic acid, methyl ester 119,407 000112-39-0 95
Hexadecanoic acid, methyl ester 119,408 000112-39-0 86
13 20.956 3.59 Propanamide 727 000079-05-0 43 No application was reported
1,3-Dioxolane-4-methanol 4690 005464-28-8 38 No application was reported
1,2-Benzenediol, 4-[2-(methylamino)ethyl]- 36,259 000501-15-5 38 No application was reported
14 21.449 1.65 Ala-gly, trimethylsilyl ester 76,119 1000333-70-1 45 No application was reported
Benzeneethanamine, 2,5-difluoro-.b eta.,3,4-trihydroxy-N-methyl- 76,728 152434-78-1 53 No application was reported
1,3-Dioxolane-4-methanol 4690 005464-28-8 53 No application was reported
15 22.497 2.33 Methyl 5,12-octadecadienoate 139,679 1000336-43-1 42 bioactive compounds toxin 24.
9,12-Octadecadienoic acid (Z, Z)-, methyl ester 139,727 000112-63-0 38 A doubly unsaturated fatty acid 18.
9,12-Octadecadienoic acid, methyl ester 139,708 002462-85-3 97 A doubly unsaturated fatty acid18.
16 22.609 22.46 9-Octadecenoic acid, methyl ester, (E)- 141,310 001937-62-8 83 No application was reported
11-Octadecenoic acid, methyl ester 141,291 052380-33-3 95 Bioactive compound has antidiarrhoeal activity 24.
trans-13-Octadecenoic acid, methyl ester 141,314 1000333-61-3 91 fatty acid has a role as a human metabolite 18.
17 22.912 6.90 Methyl stearate 143,126 000112-61-8 99 fatty acid 18.
Heptadecanoic acid, 16-methyl-, me thyl ester 143,185 005129-61-3 99 methyl-branched fatty acid 18.
Methyl stearate 143,130 000112-61-8 74 fatty acid has a role as metabolite 18.
18 23.154 7.40 Epinephrine 48,264 006539-57-7 38 a hormone and a neurotransmitter 25.
1,2-Benzenediol, 4-(2-amino-1-hydroxypropyl) 48,264 006539-57-7 32 a derivative of norepinephrine that constricts blood vessels 18,25.
1,8-Octanediamine, N,N’-dimethyl- 39,684 033563-54-1 38 No application was reported
19 25.188 1.79 1,3-Dioxolane-4-methanol 4690 005464-28-8 47 No application was reported
3-Amino-2-ethyl-butyric acid 13,898 121006-12-0 35 Bioactive compound used to determine the volatile fatty acids 18.
Metaraminol 36,214 000054-49-9 43 prevention and management of hypotension, or low blood pressure, especially when it arises as an anesthetic side effect 25.
20 27.266 2.16 Ala-gly, trimethylsilyl ester 76,119 1000333-70-1 38 No application was reported
12-Methylaminododecanoic acid, t-butyl ester 132,029 1000194-27-6 35 No application was reported
dl-3-Aminoisobutyric acid, N-methy l-, methyl ester 13,921 1000332-87-9 35 No application was reported
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The peaks in Fig. 2 indicate each biomolecule in the crude extract. The GC–MS data of the twenty biomolecules in the crude material are presented in Table 3. The peaks with the highest abundance were identified as fatty acids.

Table 3 illustrates that the lyophilized powder of the Strept lieno culture supernatant comprises several significant metabolites employed in the production of valuable pharmaceuticals18–21. This study aimed to assess the impact of laser irradiation after 10 days of incubation on the metabolism of Strept. lieno by analyzing extracellular metabolite content; furthermore, genetic alterations in Strept. lieno may be investigated in subsequent research. This study identified novel products, including thiophene. The findings demonstrated a complete reversal of the activity of the non-antibacterial Strept. lieno strain following irradiation at a wavelength of 650 nm for 2 min, which displayed a significant bactericidal impact post-irradiation. GC-MS was conducted to identify the biomolecules generated after 650 nm irradiation, with findings presented in Table 4; Fig. 3.

The curve shape distinctly transitions from the 600–1200 s⁻¹ range (of non-irradiated Streptomyces lieno, exhibiting a light pink hue) to the 2200–2800 s⁻¹ range, where the irradiated strain culture displays a dark brown coloration.

Table 4 prominently displays numerous significant bioactive chemicals, including thiophene derivatives, hormones and neurotransmitters such as; epinephrine, derivatives of norepinephrine, vitamin B6 precursors, toxins, hydrogen bond donors, anti-hypertension reagents and so on.

Discussion

The local Strept.lieno isolate examined in this study exhibited minimal antibacterial activity when compared to the antibacterial efficacy of Strept thin; specifically, Strept.lieno produced a narrow inhibition zone with a diameter of less than 5 mm in a sensitivity test against S. aureus and showed no effect on E. coli. The GC-Mass data (Table 3) indicated the absence of antibiotic compounds.

In the preliminary study, the isolate with notable antibacterial potential, Strept thin, showed a slight increase in activity post-laser irradiation, while Strept.lieno, which exhibited reduced inhibitory effects, displayed significant efficacy in the production of antimicrobial compounds following laser irradiation.The Strept lieno groups irradiated at a wavelength of 650 nm exhibited a significant enhancement in antibacterial activity. In the second group, the Strept lieno subjected to 532 nm irradiation were likewise impacted, albeit to a lesser degree than those exposed to the red wavelength.

In general, inducing changes in biological components requires the absorption of laser photon energy by specific biomolecules, such as cytochrome c oxidase, acetyl carrier protein and acetyl-CoA. The precursors for fatty acid biosynthesis are derived from the acetyl-CoA pool9. This energy stimulates several photochemical reactions and changes the redox potential inside bacterial cells26. If energy is transferred to biomaterials via electron excitation, this leads to the formation of covalent bonds and the release of reactive oxygen species (ROS). Consequently, these ROS cause direct damage to bacterial enzymes, denature proteins in some organelles, and increase the synthesis of antioxidants, including oxidoreductases, as a defence mechanism to protect RNA from total damage27.

According to our findings in this work, irradiation of the Strept lieno isolates at 650 nm led to the production of more antibacterial molecules that irradiation at 532 nm did.

The response to the 532 nm wavelength may arise from the presence of various chromophores in the cytoplasm of cells, particularly those containing haem groups, such as protoporphyrin IX, which demonstrates significant absorption of green light. This absorption leads to mainly the transduction and amplification of photosignals, which is maintained after 10 days of incubation of the bacterial culture28. On the other hand, many studies have reported that red and infrared laser light is absorbed mainly by cytochrome c oxidase and that water molecules in the cytoplasm stimulate redox reactions by releasing reactive oxygen species (ROS), such as H2O2, OH−, and NO owing to photodissociation from cytochrome c oxidase29,30. In this research, the local Strept. lieno isolate began producing thiophene only after 650 nm irradiation, as this compound was not detected by GC–MS and this isolate was not active in the sensitivity test before irradiation. GC–MS provided information about the components in the supernatant, which may help us to understand how laser light stimulates the biosynthesis of thiophene and possibly its precursors.

Figure 2 shows the compounds identified by GC–MS according to spectra in the NIST database from the control Strept. lieno sample (Fig. 3) and the Strept. lieno sample irradiated at a wavelength of 650 nm nm for 2 min.

Figures 2 and 3 show that both curves have the same number of peaks, which represent the biomolecules released into the fermentation broth supernatant of Strept. However, in Fig. 3, the values for some peaks changed markedly; for example, the first peak at 5.137 in Fig. 2 appeared at 5.163 in Fig. 3, which indicates that the structures of the compounds changed by consecutive photochemical reactions in the bacterial cells. To identify the compounds released before and after laser irradiation, the presence of thiophene was investigated (Table 2).

Thiophene, also known as acetylenic thiophene, is biosynthetically derived from fatty acids or polyacetylenes through acetylene intermediates. Thiophenes are a class of sulfur-containing molecules usually composed of one to five thiophene units and various alkyl groups on the α-carbon of the terminal ring31.

Therefore, fatty acids and polyacetylenes are derived from oxidative stress upon laser irradiation and cause direct DNA damage through the formation of photoproducts, including cyclobutane‒pyrimidine dimers and pyrimidine-(6‒4)-pyrimidone32–35. These dimers eventually form the fatty acid polyacetylene, which is a precursor for thiophene. This hypothesis was created on the basis of the abundance of cyclobutane compounds in the supernatants of the irradiated samples (Table 2).

Conclusion

Laser light can induce certain natural isolates of Strept. to synthesize thiophenes, which, due to their bioactivities, have garnered significant interest from researchers for further exploration of their mechanisms, efficacy, and safety. The structural diversity of thiophenes may hold significant synthetic value as innovative chemical entities for drug discovery. This study may stimulate additional research on the appropriate dosage and wavelength of laser irradiation for related investigations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary Material 1 (77.6KB, docx)

Author contributions

N.J planned and wrote the paperT.A isolate and characterized the Streptomycetes Both authors reviewed the paper.

Funding

According to article (11/sixth) of the revised University Service Law (23 of 2008), the University of Baghdad is providing conditional financial funding for the current study.

Data availability

No datasets were generated or analysed during the current study.

Declarations

Competing interests

The authors declare no competing interests.

Ethical approval

Not applicable.

Clinical trial number

Not applicable.

Footnotes

Publisher’s note

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Supplementary Materials

Supplementary Material 1 (77.6KB, docx)

Data Availability Statement

No datasets were generated or analysed during the current study.

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