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An Investigation of Glyceollin I's Inhibitory Effect on The Mammalian Adenylyl (글리세올린 I의 아데니닐 고리화 효소 활성 억제 효능과 결합 부위 비교 분석)

  • Kim, Dong-Chan;Kim, Nam Doo;Kim, Sung In;Jang, Chul-Soo;Kweon, Chang Oh;Kim, Byung Weon;Ryu, Jae-Ki;Kim, Hyun-Kyung;Lee, Suk Jun;Lee, Seungho;Kim, Dongjin
    • Journal of Life Science
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    • v.23 no.5
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    • pp.609-615
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    • 2013
  • Glyceollin I has gained attention as a useful therapy for various dermatological diseases. However, the binding property of glyceollin I to the mammalian adenylyl cyclase (hereafter mAC), a critical target enzyme for the down-regulation of skin melanogenesis, has not been fully explored. To clarify the action mechanism between glyceollin I and mAC, we first investigated the molecular docking property of glyceollin I to mAC and compared with that of SQ22,536, a well-known mAC inhibitor, to mAC. Glyceollin I showed superiority by forming three hydrogen bonds with Asp 1018, Trp 1020, and Asn 1025, which exist in the catalytic site of mAC. However, SQ22,536 formed only two hydrogen bonds with Asp 1018 and Asn 1025. Secondly, we confirmed that glyceollin I effectively inhibits the formation of forskolin-induced cAMP and the phosphorylation of PKA from a cell-based assay. Long term treatment with glyceollin I had little effect on the cell viability. The findings of the present study also suggest that glyceollin I may be extended to be used as an effective inhibitor of hyperpigmentation.

The Inhibitory Effects of Pogostemon cablin Bentham Extract on Melanogenesis (광곽향 추출물의 멜라닌 생성 저해 효과)

  • Bae, Seong-Yun;Lee, Eung-Ji;Son, Rak-Ho;Lee, Yong-Hwa
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.1
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    • pp.33-39
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    • 2009
  • To develop a new natural whitening agent for cosmetics, we investigated the inhibitory effects of Pogostemon cablin Bentham extracts (PCE) and its active component on melanogenesis. PCE showed ROS scavenging activities in 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and xanthine/xanthine oxidase system with the $IC_{50}$ values of $24.2{\pm}2.85{\mu}g/mL$ and $IC_{50}=118{\pm}0.43{\mu}g/mL$, respectively. PCE reduced melanin contents of B16 melanoma cells in a dose-dependant manner and decreased to about 23 % at a concentration of $20{\mu}g/mL$ without cell cytotoxicity (below $100{\mu}g/mL$). And the PCE reduced intracellular tyrosinase activity about 18 % at concentration of $50{\mu}g/mL$. We purified one active compound from PCE and identified its structure. It was identified as patchouli alcohol, sesquiterpene family, by 1H-NMR, $13_C$-NMR, and Mass analysis. Patchouli alcohol also inhibited ROS scavenging activities in DPPH radical and xanthine/xanthine oxidase system with the $IC_{50}$ values of $3.14{\pm}0.12{\mu}g/mL$ and $49{\pm}3.24{\mu}g/mL$, respectively. Patchouli alcohol inhibited melanin synthesis in a dose dependent manner ($IC_{50}=3.9{\mu}g/mL$). And the patchouli alcohol reduced intracellular tyrosinase activity about 40 % at concentration of $10{\mu}g/mL$. Patchouli alcohol inhibited tyrosinase and TRP-2 expression at protein level. These results suggest that PCE and patchouli alcohol reduced melanin formation by the inhibited of tyrosinase activity and expression in B16 melanoma cells. Therefore, we suggest that PCE could be used as a useful whitening agent.

Melanogenesis regulatory constituents from Premna serratifolia wood collected in Myanmar

  • WOO, SO-YEUN
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.21-22
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    • 2019
  • Melanin is a mixture of pigmented biopolymers synthesized by epidermal melanocytes that determine the skin, eye, and hair colors. Melanocytes produce two different kinds of melanin, eumelanin (dark brown/black insoluble pigments found in dark skin and dark hair and pheomelanin (lighter red/yellow). The biological role of melanin is to prevent skin damage by ultraviolet (UV) radiation. However, the overproduction or deficiency of melanin synthesis could lead to serious dermatological problems, which include melasma, melanoderma, lentigo, and vitiligo. Therefore, regulating melanin production is important to prevent the pigmentation disorders. Myanmar has a rich in natural resources. However, the chemical constituents of these natural resources in Myanmar have not been fully investigated. In the effort to search for compounds with anti-melanin deposition activity from Myanmar natural resources, five plants were collected in Myanmar. Extracts of these collected five plants were tested for anti-melanin deposition activity against a mouse melanoma cell line (B16-F10) induced with ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) and 3-isobutyl-1-methylxanthine (IBMX), and their anti-melanin deposition activities were compared with the positive control, arbutin. Among the tested extracts, the CHCl3 extracts of the Premna serratifolia (syn: P. integrifolia) wood showed anti-melanin deposition activities with IC50 values of $81.3{\mu}g/mL$. Hence, this study aims to identify secondary metabolites with anti-melanin deposition activity from P. serratifolia wood of Myanmar. P. serratifolia belongs to the Verbenaceae family and is widely distributed in near western sea coast from South Asia to South East Asia, which include India, Malaysia, Vietnam, Cambodia, and Sri Lanka. People in Tanintharyi region located in the southern part of Myanmar utilize the P. serratifolia, Sperethusa crenulata, Naringi crenulata, and Limonia acidissima as Thanaka, traditional cosmetics in Myanmar. Thanaka is applied in the form of paste onto skins to make it smooth and clear, as well as to prevent wrinkles, skin aging, excessive facial oil, pimples, blackheads, and whiteheads. However, the chemical constituents responsible for their cosmetic properties are yet to be identified. Moreover, the chemical constituents of P. serratifolia was almost uncharacterized. Investigation of the P. serratifolia chemical constituents is thus an attractive endeavor to discover new anti-melanin deposition active compounds. The investigation of the chemical constituents of the active CHCl3 extract of P. serratifolia led to isolation of four new lignoids, premnan A (1), premnan B (2), taungtangyiol C (3), and 7,9-dihydroxydolichanthin B (4), together with premnan C (5) (assumed to be an artifact), one natural newlignoid,(3R,4S)-4-(1,3-benzodioxol-5-ylcarbonyl)-3-[(R)-1-(1,3-benzo dioxol-5-yl)-1-hydroxy methyl]tetrahydro-2-furanone (6), and five known compounds (7-11)1,2). The structures of all isolated compounds were determined on the basis of their spectroscopic data and by comparison with the reported literatures. The absolute configurations of 1-3 and 5 were also determined by optical rotation and circular dichroism (CD) data analyses1). The anti-melanin deposition activities of all the isolated compounds were evaluated against B16-F10 cell line. 7,9-Dihydroxydolichanthin B (4) and ($2{\alpha},3{\alpha}$)-olean-12-en-28-oic acid (11) showed strong anti-melanin deposition activities with IC50 values of 18.4 and $11.2{\mu}M$, respectively, without cytotoxicity2). On the other hand, compounds 1-3, 5, and 7 showed melanogenesis enhancing activities1). To better understand their anti-melanin deposition mechanism, the effects of 4 and 11 on tyrosinase activities were investigated. The assay indicated that compounds 4 and 11 did not inhibit tyrosinase. Furthermore, we also examined the mRNA expression of microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), tyrosinase-related protein-1 (TRP-1), and tyrosinase-related protein-2 (TRP-2). Compounds 4 and 11 down-regulated the expression of Tyr and Mitf mRNAs, respectively. Although the P. serratifolia wood has been used as traditional cosmetics in Myanmar for centuries, there are no scientific evidences to support its effectiveness as cosmetics. Investigation of the anti-melanin deposition activity of the chemical constituents of P. serratifolia thus provided insight into the effectiveness of the P. serratifolia wood as a cosmetic agent.

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Association of β3-Adrenergic Receptor Polymorphisms and High-Density Lipoprotein Cholesterol (고밀도 지단백 콜레스테롤과 베타 3-아드레날린성 수용체 유전자 변이와의 관련성)

  • Yu, Byeng-Chul;Jeon, Man-Joong;Lee, Yong-Hwan
    • Journal of Life Science
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    • v.19 no.5
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    • pp.664-670
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    • 2009
  • The ${\beta}$3-adrenergic receptor (ADRB3) is expressed mainly in visceral adipose tissue and is thought to contribute to lipolysis and the delivery of free fatty acids to the portal vein. This study was aimed at evaluating the association between high-density lipoprotein cholesterol (HDL-C) and ADRB3 genetic polymophisms. A total of 991 healthy examinees who were examined in a university hospital, located in Busan City, between May and December 2006 were enrolled in this study. Height, weight, body mass index, waist circumference, and systolic and diastolic blood pressures of the subjects were examined. Intravenous concentrations of fasting blood glucose, total cholesterol, HDL-C, low-density lipoprotein cholesterol, and triglyceride were also measured. After extracting DNA from the subjects, mutations of the +188T>C (Trp64Arg) of exon 1 and +3893T>C of intron 2 on the ADRB3 gene were genotyped using the single base extension method. We have identified a novel mutation of ADRB3 that is located in intron 2. The frequency of its minor allele was 0.164. Both the +188T>C mutation of exon 1 and +3893T>C mutation of intron 2 were significantly associated with HDL-C. The mean concentration of serum HDL-C was significantly lower in the presence of their minor allele 'C'. These results suggest that both mutations of +188T>C of exon 1 and +3895T>C of intron 2 have significant associations with HDL-C in the Korean population.

Effects of Low Crude Protein Diets Supplemented with Synthetic Amino Acids on Performance, Nutrient Utilization and Carcass Characteristics in Finishing Pigs Reared Using a Phase Feeding Regimen

  • Lee, J.H.;Kim, J.H.;Kim, J.D.;Kim, S.W.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.5
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    • pp.655-667
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    • 2001
  • This experiment was conducted to investigate the effect of feeding a low CP diet supplemented with synthetic amino acids on performance, nutrient utilization and carcass characteristics of finishing pigs fed under a three-phase feeding regimen. Ninety-six finishing pigs (Landrace$\times$Large White$\times$Duroc), $55.75kg{\pm}0.65$ of initial body weight, were blocked by weight and sex and allotted to four dietary treatments in a randomized block design. There were six pens per treatment and four pigs per pen. Pigs were fed a 16%-14%-12% CP (for phase I-II-III, respectively), sequence of diets. Dietary treatments were 1) Control, 2) Con+L (a sequence of diets reduced in CP by l percentage unit with lysine (L) supplementation, 3) Con+LMT (a sequence of diets reduced in CP by 2 percentage unit with LYS, methionine (MET) and threonine (THE) supplementation) and 4) Con+LMTT (a sequence of diets reduced in CP by 3 percentage unit with LYS, MET, THR and tryptophan (TRP) supplementation). The finishing period (55 to 105 kg) was divided into three phases (55 to 72 kg, 72 to 90 kg and 90 to 105 kg). Pigs fed either the control or Con+L diet grew faster (p<0.05) than pigs fed the Con-LMT or Con+LMTT diet. There was no difference in ADFI among dietary treatments. Phosphorus (P) digestibility was lowest in the control group and highest in the Con+LMTT group (p<0.05). Within each phase, no significant differences in dry matter (DM) and CP digestibilities were found. Although some amino acid digestibilities were affected by dietary treatments, digestibilities of essential amino acids (EAA), non-essential amino acids (NEAA) and total amino acid were not significantly influenced by dietary treatments. For the entire experiment periods, Con+L, Con+LMT and Con+LMTT treatments resulted in 13.4, 18.8 and 21.6% lower total N excretion compared with the control. Con+LMT and Con+LMTT treatments showed significantly lower BUN concentration compared with the control and Con+L treatment (p<0.05), but there was no significant difference in BUN concentration between pigs fed the control and Con+L treatment or between pigs fed Con+LMT and Con+LMTT treatments (p>0.05). Carcass length, backfat thickness and carcass grade were not significantly affected by dietary treatments (p>0.05). In conclusion, reducing dietary CP level by 1 percentage unit and supplementing only LYS at each phase could be a very beneficial feeding strategy for finishing pigs fed under a three phase feeding regimen in terms of both environmental and economical aspects.

Ileal Digestibility of Amino Acids in Conventional and Low-Kunitz Soybean Products Fed to Weanling Pigs

  • Goebel, K.P.;Stein, H.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.1
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    • pp.88-95
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    • 2011
  • An experiment was conducted to determine the standardized ileal digestibility (SID) of amino acids (AA) in four sources of full-fat soybeans (FFSB) and in one source of soybean meal (SBM). The FFSB had different concentrations of trypsin inhibitor units (TIU) and included two sources of conventional FFSB, and two sources of a soybean variety that was selected for a reduced concentration of the Kunitz trypsin inhibitor. The conventional FFSB was either low temperature-processed (LT-FFSB-CV; 37.7% CP, 35.4 TIU/mg) or high temperature-processed (HT-FFSB-CV; 40.5% CP, 4.4 TIU/mg). The low-Kunitz FFSB was also either low temperature-processed (LT-FFSB-LK; 36.2% CP, 23.5 TIU/mg) or high temperature-processed HT-FFSB-LK; (38.2% CP, 4.0 TIU/mg). The SBM contained 47.5% CP and 3.20 TIU/mg. Twelve weanling barrows (initial BW: $11.1{\pm}1.3\;kg$) were fitted with a T-cannula in the distal ileum. Pigs were allotted to a replicated $6{\times}6$ Latin square design with six diets and six periods per square. Five diets were prepared using each of the soybean sources as the only source of AA in the diet. An N-free diet was also included in the experiment to measure basal endogenous losses of AA. The two low temperature-processed FFSB had lower (p<0.05) AID and SID values for all indispensable AA than the two high temperature-processed FFSB and SBM. The SID values for all indispensible AA except Trp were greater (p<0.05) in LT-FFSB-LK than in LT-FFSB-CV, but the SID of AA in HT-FFSB-CV and HT-FFSB-LK were not different. The SID of AA in SBM were not different from the SID in HT-FFSB-CV and in HT-FFSB-LK. Results of this experiment show that a reduction of the TIU from 35.4 to 23.5 TIU/mg will improve the SID of AA, but this reduction is not sufficient to completely ameliorate the negative impact of trypsin inhibitors. Results also show that the SID of AA in high temperature-processed FFSB is similar to that in de-hulled SBM.

Interaction of Barley Acetolactate Synthase with Triazolopyrimidine Inhibitors (Triazolopyrimidine계 저해제와 보리 Acetolactate Synthase와의 상호작용)

  • Lee, Jae Soeb;Chang, Soo Ik;Nam Goong, Sung Keon;Shin, Jung Hyu;Choi, Jung Do
    • Journal of the Korean Chemical Society
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    • v.42 no.3
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    • pp.306-314
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    • 1998
  • Acetolactate synthase (ALS) is the common enzyme in the biosynthesis of branched chain amino acids, Val, Leu, and Ile in bacteria, yeast, and higher plants. The enzyme is target site of several classes of structually diverse herbicides, including the sulfonylureas, the imidazolinones, the triazolopyrimidines, and the primidyl-oxy-benzoates. We have synthesized new triazolopyrimidine (TP) derivatives, and determined their inhibitory activities on barley ALS. $lC_{50}$ values for the active compounds were 3.2 nM-0.62 mM, and some of them appeared to be potent inhibitors. The progress curves for inhibition of ALS by TP4, a representative derivative, indicated that the extent of inhibition increased with incubation time. The inhibition of ALS by TP4 showed mixed-type inhibition with respect to pyruvate. Dual inhibition analyses of TP4 versus imidazolinone Cadre and feedback inhibitor Leu suggested that three different classes of inhibitors bind to ALS in a mutually exclusive manner. Chemical modification of tyrosyl residues of ALS decreased sensitivity of ALS to TP4, while modification of tryptophan and cysteine did not affect the sensitivity.

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Characterization and application of the proteins isolated from edible insects (식용곤충 분리단백의 특성 및 활용연구)

  • Kim, Sun-Min;An, Chae-Won;Han, Jung-Ah
    • Korean Journal of Food Science and Technology
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    • v.51 no.6
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    • pp.537-542
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    • 2019
  • Proteins were isolated from four species of edible insects, Bombyx mori, B; Tenebrio molitor, T; Protaetia brevitarsis, P; and Gryllus bimaculatus, G, and their properties were analyzed and compared with those of whey proteins. The yield of the protein isolated from raw materials was 65.0-75.0%. Among three branched chain amino acids (BCAA), the amounts of valine in all the insects were similar to that of the whey protein. The BCAA content was 16.8 and 16.4% in G and T, respectively. The total polyphenolic content and the antioxidant activity of the insect proteins were significantly higher than those of the whey proteins. Based on the sensory test of the protein powder, the whey protein could be replaced by 25% of the isolated protein from the insects, but the taste and flavor needs to be improved. Although the BCAA content of insect proteins was lower than that of the whey protein (by 73-76%), their application could be useful because of high polyphenolic content and the anti-oxidative activity.

Effects of amino acid composition in pig diet on odorous compounds and microbial characteristics of swine excreta

  • Recharla, Neeraja;Kim, Kihyun;Park, Juncheol;Jeong, Jinyoung;Jeong, Yongdae;Lee, Hyunjeong;Hwang, Okhwa;Ryu, Jaehyoung;Baek, Youlchang;Oh, Youngkyun;Park, Sungkwon
    • Journal of Animal Science and Technology
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    • v.59 no.12
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    • pp.28.1-28.8
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    • 2017
  • Background: Major amino acids in pig diets are Lys, Met, Thr, and Trp, but little is known about the requirements for the other essential amino acids, especially on odorous compounds and microbial characteristics in feces of growing-finishing pigs. To this end, different levels of amino acid composition added to diets to investigate the effects of amino acid composition on microbial characteristics and odorous compounds concentration. Methods: A total eight (n = 8) barrows (Landrace ${\times}$ Yorkshire ${\times}$ Duroc) with an average bodyweight of $89.38{\pm}3.3kg$ were individually fed diets formulated by Korean Feeding Standards 2007 (old version) or 2012 (updated with ideal protein concept) in metabolism crates with two replication. After 15-day adaptation period, fresh faecal samples were collected directly from pigs every week for 4 weeks and analysed for total volatile fatty acids (VFA), phenols and indoles by using gas chromatography. The nitrogen was determined by Kjeldahl method. Bacterial communities were detected by using a 454 FLX titanium pyrosequencing system. Results: Level of VFA tended to be greater in 2012 than 2007 group. Among VFAs, 2012 group had greater (p < 0.05) level of short chain fatty acids (SCFA) than control.Concentration of odorous compounds in feces was also affected by amino acid composition in pig diet. Levels of ammonium and indoles tended to be higher in 2012 group when compared with 2007 group.Concentration of phenols, p-cresol, biochemical oxygen demand, and total Kjeldahl nitrogen, however, were lower (P < 0.05) in 2012 treatment group compare to 2007. The proportion of Firmicute phylum were decreased, while the Bacteriodetes phylum proportion increased and bacterial genera includingCoprococcus, Bacillus, and Bacteroides increased (p < 0.05) in 2012 compare to 2007 group. Conclusion: Results from our current study indicates that well balanced amino acid composition reduces odor by modulating the gut microbial community. Administration of pig diet formulated with the ideal protein concept may help improve gut fermentation as well as reduce the odor causing compounds in pig manure.

Mutations in Streptomycin Resistance Genes and Their Relationship to Streptomycin Resistance and Lineage of Mycobacterium tuberculosis Thai Isolates

  • Hlaing, Yin Moe;Tongtawe, Pongsri;Tapchaisri, Pramuan;Thanongsaksrikul, Jeeraphong;Thawornwan, Unchana;Archanachan, Buppa;Srimanote, Potjanee
    • Tuberculosis and Respiratory Diseases
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    • v.80 no.2
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    • pp.159-168
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    • 2017
  • Background: Streptomycin (SM) is recommended by the World Health Organization (WHO) as a part of standard regimens for retreating multidrug-resistant tuberculosis (MDR-TB) cases. The incidence of MDR-TB in retreatment cases was 19% in Thailand. To date, information on SM resistance (SMR) gene mutations correlated to the SMR of Mycobacterium tuberculosis Thai isolates is limited. In this study, the mutations in rpsL, rrs, gidB, and whiB7 were investigated and their association to SMR and the lineage of M. tuberculosis were explored. Methods: The lineages of 287 M. tuberculosis collected from 2007 to 2011 were identified by spoligotyping. Drug susceptibility profiles were evaluated by the absolute concentration method. Mutations in SMR genes of 46 SM-resistant and 55 SM-susceptible isolates were examined by DNA sequencing. Results: Three rpsL (Lys43Arg, Lys88Arg, and Lys88Thr) and two gidB (Trp45Ter and Gly69Asp) mutations were present exclusively in the SM resistant M. tuberculosis. Lys43Arg rpsL was the most predominant SMR mutations (69.6%) and prevailed among Beijing isolates (p<0.001). No SMR-related mutation in was found rrs. The combination of rpsL and gidB mutations provided 76.1% sensitivity for detecting SMR in M. tuberculosis Thai isolates. whiB7 was not responsible for SMR in SM resistant isolates lacking rpsL and rrs mutations. The significance of the three gidB mutations, 276A>C, 615A>G, and 330G>T, as lineage signatures for Beijing and EAI were underscored. This study identified 423G>A gidB as a novel sub-lineage marker for EAI6-BGD1. Conclusion: Our study suggested that the majority of SMR in M. tuberculosis Thai isolates were responsible by rpsL and gidB polymorphisms constantly providing the novel lineage specific makers.