Hypocrellin A (HA), a photodynamic perylenequinone pigment from Shiraia fruiting bodies, functions as an efficient photosensitizer for clinical photodynamic therapy. Glucose-6-phosphate dehydrogenase (G6PDH), the rate-limiting enzyme of the pentose phosphate pathway (PPP), governs carbon flux into NADPH production. This study elucidates G6PDH’s regulatory role in HA biosynthesis in Shiraia sp. S9. Bamboo polysaccharide (BPS) elicitation (100 mg/L) significantly enhanced HA production to 428.1 mg/L, 1.6-fold higher than controls after 5 days. We cloned the G6PDH gene and demonstrated that BPS upregulated its expression and activity, concomitant with increased reactive oxygen species (ROS; H2O2 and O2•−) and nitric oxide (NO) generation. ROS production was mediated by NADPH oxidase induction, while NO generation was attributed to elevated nitric oxide synthase and nitrate reductase activities. Critically, the G6PDH inhibitor glucosamine (1.0 mM) suppressed both H2O2 and NO production. These ROS/NO signals upregulated key HA biosynthetic (PKS, Omef) and transport (MFS) genes. Our findings establish G6PDH as a central regulator of BPS-induced HA biosynthesis via ROS/NO signaling, revealing novel metabolic crosstalk between the PPP and fungal perylenequinone biosynthesis. This work presents BPS elicitation as a biotechnological strategy for scalable HA production in Shiraia mycelium cultures.

BackgroundHepatocellular carcinoma (HCC) is a highly prevalent malignant tumor worldwide. Evidence showed that polyamine metabolism plays a crucial part in the regulation of cancer onset and development, however, its clinical significance in HCC remains unclear.MethodsBulk RNA sequencing (RNA-seq) and single-cell RNA sequencing (scRNA-seq) data of HCC were collected from public databases. Polyamine metabolism-related genes (PMRGs) were obtained from the MSigDB database. The molecular subtypes of HCC were classified by ConsensusClusterPlus package, and differentially expressed genes (DEGs) of the molecular subtypes were identified by the limma package, followed by enrichment analysis with clusterProfiler package. Univariate Cox and Lasso Cox regression analyses were performed to screen core genes, construct risk model, and develop a nomogram integrating clinical characteristics for survival prediction. The obtained biomarkers were validated using in vitro experiments (CCK8, wound healing, and Transwell assay). The Tumor Immune Estimation Resource (TIMER), MCP-counter, and Cell Identification by Estimating Relative Subsets of RNA Transcripts (CIBERSORT) methods were employed for immune cell infiltration analysis. Finally, drug sensitivity of the HCC samples was analyzed with the oncoPredict package.ResultsThis study identified two molecular subtypes (C1 and C2), with C2 demonstrating a more favorable prognosis. Glucose-6-phosphate dehydrogenase (G6PD), alcohol dehydrogenase 4 (ADH4), S100 calcium binding protein A9 (S100A9), aldo-keto reductase family 1 member B15 (AKR1B15) were predicted as the biomarkers for HCC. Cell experiment results showed that the expressions of G6PD, AKR1B15, and S100A9 were all notably elevated in HuH-7 cells. Moreover, the loss of G6PD gene expression reduced the viability, migratory, and invasive capabilities of HCC cells. Patients with a high RiskScore had a lower survival rate than those with a low RiskScore. Scores of immune cells such as Tregs and M0 macrophages were higher in the high-risk group, and 13 drugs were found to be significantly linked to the RiskScore. Single-cell analysis showed that G6PD and S100A9 were high-expressed mainly in hematopoietic progenitor cells (HPCs) and macrophages.ConclusionIn conclusion, this study screened four key genes based on PMRGs and constructed a risk model to effectively predict the prognosis of HCC, providing novel potential targets and theoretical basis for the molecular subtyping and individualized treatment of HCC.

Hypoxia and activation of the pentose phosphate pathway (PPP), as well as overexpression of glucose-6-phosphate dehydrogenase (G6PD), are hallmark features of glioblastomas (GBM), contributing significantly to tumor progression metabolic adaptation and drug resistance. This study aimed to evaluate the cytotoxic effects of nine synthetic compounds incorporating annulated benzimidazole and nitrothiazole scaffolds in two glioblastoma cell lines (A172 and U87-MG) under both normoxic and hypoxic conditions. Three compounds (BZM-7, BZM-9, and CNZ-3) demonstrated potent anticancer activity, with CNZ-3 exhibiting the highest efficacy, particularly in hypoxia. The study further investigated the effects of these compounds on the expression of the G6PD gene, as well as post-translational regulatory genes SIRT2 and KAT9, and the angiogenesis-related VEGF gene. Transcriptional analyses showed that the nitrothiazole-derived compound CNZ-3 significantly downregulated G6PD, SIRT2, KAT9 and VEGF expression under hypoxic conditions, suggesting selective interference with hypoxia-adaptative pathways. In contrast, BZM-7 and BZM-9 showed distinct expression patterns, indicating diverse mechanisms of action despite structural similarity. In addition, BZM-7, BZM-9, and CNZ-3 were identified as potent inhibitors of recombinant G6PD, demonstrating both enzymatic inhibition and structural alterations, suggesting that G6PD could be a relevant therapeutic target for these compounds. Furthermore, molecular docking analysis revealed favorable binding interactions between the compounds and key amino acids of the G6PD, reinforcing their potential as a direct enzyme inhibitors. These findings highlight the pivotal role of G6PD in gliomas under hypoxic conditions and support its inhibition as a promising therapeutic strategy.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a widespread enzymopathy affecting approximately 500 million individuals that represents a significant global health issue. Among the more than 230 identified mutations in the G6PD gene, six class A variants—G6PD Utrecht (Pro409Ser), G6PD Suwalki (Pro409Arg), G6PD Merlo (Pro409Gln), G6PD Kawasaki (Gly410Ala), G6PD Shinagawa (Gly410Asp), and G6PD Riverside (Gly410Cys)—are located in the beta-loop near the NADP+ binding site. These mutations are of particular interest due to their association with severe hematologic phenotypes, including chronic hemolytic anemia, as well as their proposed role in the allosteric regulation of G6PD multimerization. This study presents a comprehensive biochemical and functional characterization of these clinically relevant G6PD variants. The variant enzymes were cloned, expressed, and purified for characterization. Kinetic parameters and thermal stability assays, complemented by molecular dynamics simulations (MDS), were employed to elucidate the structural impacts of the mutations. Our results demonstrate that these mutations significantly impair protein function, characterized by reduced affinity for glucose-6-phosphate (G6P) and NADP+, as well as altered thermal stability compared with wild-type G6PD. MDS revealed that point mutations in the βN- and βM-sheets in the NADP+s region propagate subtle conformational changes, ultimately affecting the NADP+c region and the G6P binding cavity. Furthermore, secondary structure element analyses of the simulation data showed that Pro409 and Gly410 point mutations propagate several changes around residues 195–210 (G6P binding site) and 380–400 (NADP+s), explaining their effect on overall catalytic performance. These findings enhance our understanding of the molecular mechanisms underlying G6PD deficiency and its clinical implications, providing a foundation for future therapeutic strategies aimed at mitigating the effects of these variants.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most prevalent enzyme deficiency of human erythrocytes, affecting more than 500 million people globally. G6PD deficiency is the most com- mon hemolytic X-linked genetic disease, which shows a higher prevalence rate in the malaria-endemic population. This review essay aims to examine the overall frequency of G6PD deficiency in the Indian population using systemic review, with a special emphasis on the differential prevalence rate across populations. PubMed, Google Scholar, and ScienceDirect databases were searched to examine the scientific studies focusing on the prevalence of G6PD deficiency among Indian populations. A total of 32 studies were selected after following several inclusion-exclusion criteria, which comprised a total of 115,300 participants. It is observed that the G6PD deficiency is higher among males than females, which is expected as this enzymopathy is caused by inherited mutations of the X-linked gene G6PD. Moreover, the disorder is considerably greater among populations that prefer consanguineal and endogamous marriages, such as Parsi, Muslim, and tribal populations, as compared with nontribal and caste populations. Hence, culturally based marriage practices may help to explain differences in the frequency of this hemolytic genetic disorder among Indian populations.

Plasmodium vivax malaria remains a significant global health challenge, complicated by the parasite’s ability to form dormant liver stages (hypnozoites) that cause relapses. Radical cure of P. vivax malaria requires administration of a hypnozoitocidal drug, such as primaquine or tafenoquine. However, these drugs can cause severe haemolysis in individuals with glucose-6-phosphate dehydrogenase (G6PD) deficiency. G6PD deficiency is caused by more than 230 different variants at the gene level that confer different degrees of deficiency phenotypically. Understanding the distribution of different G6PD variants in affected populations is essential to inform safer antimalarial treatment strategies. This study aimed to develop a cost-effective sequencing assay targeting key regions of the G6PD gene, suitable for field deployment. A novel assay based on Nanopore technology was designed to amplify two amplicons covering exon 3 to exon 13, focusing on known variants associated with enzyme deficiency. A total of 79 samples from individuals in Cambodia, Vietnam, Afghanistan, and China were sequenced, and a bioinformatics pipeline was created for the targeted variant calling of 192 G6PD SNP mutations. The assay demonstrated reliable detection of known variants, with high concordance between runs, within runs, and with Sanger sequencing.The Nanopore MinION long-amplicon sequencing assay offers a robust and portable solution for large-scale G6PD genotyping in low-resource settings, that will improve malaria control and elimination strategies by enabling safer antimalarial treatment.

Glucose-6-phosphate dehydrogenase (G6PDH) is a critical enzyme in the pentose phosphate pathway, playing an essential role in plant growth, development, and adaptation to abiotic stress. In this study, we identified four members of the G6PDH gene family in the ‘Zunla-1’ genome, designating them as CaG6PDH1-CaG6PDH4. Multiple sequence alignment revealed that the four protein sequences of pepper contain three unique binding sites characteristic of G6PDH: the substrate binding site, the NADP binding site and the Rossmann fold. The phylogenetic tree, motifs, and gene structure analysis indicate that the CaG6PDH gene sequence is relatively conserved and structurally similar, with a close relationship to the sequence of Solanaceae G6PDH members. The collinearity analysis showed that there were two pairs of collinearity between the CaG6PDH genes and the AtG6PDH genes, as well as the SiG6PDH genes. Additionally, numerous cis-elements associated with stress responses, hormone regulation, development, and light responses were identified in the promoter region of the CaG6PDH gene. Furthermore, the various members of the pepper CaG6PDH gene family exhibit specific expression patterns across different tissues and demonstrate significant variations in response to abiotic stress and phytohormone treatments, particularly the CaG6PDH1 and CaG6PDH2 genes. Subcellular localization studies indicate that CaG6PDH2 is located in chloroplasts. We conducted further investigations into the role of CaG6PDH2 in response to cold stress using Virus-Induced Gene Silencing (VIGS) technology. The tissues of seedlings with silenced CaG6PDH2 exhibited significant damage and displayed a more pronounced cold damage phenotype. This observation is further supported by the accumulation of reactive oxygen species (ROS), the activity of antioxidant enzymes, and a reduction in the expression of cold-responsive genes. In conclusion, the findings of this study indicate that CaG6PDH2 plays an important role in cold stress response and may serve as a potential gene for cultivating cold-tolerant pepper varieties.

Rapid and reliable detection of G6PD mutations using recombinase polymerase amplification coupled with lateral flow strip.

Screening G6PD Mutations in Blood Donors by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry with High-Throughput and Multiple Targets.

The data of 298 patients with auditory neuropathy (AN) who underwent genetic testing in Chinese PLA General Hospital from August 2015 to June 2024 were retrospectively reviewed, and 12 cases (4.0%) carried pathogenic glucose-6-phosphate dehydrogenase (G6PD) gene mutations were enrolled. Among them, 11 patients were from Guangxi. There were 11 cases (9 males and 2 females) that showed symptoms of G6PD deficiency, and 10 cases had neonatal hyperbilirubinemia. The audiological characteristics of the 12 patients were typical AN manifestations, with the age of onset of hearing loss of 0 (0, 1) month, and 11 cases were classified as infant AN. Genetic analysis of the 12 patients identified four G6PD mutation, with the c.1388G>A pathogenic mutation being the most prevalent (8/12). Additionally, three deafness-associated genes, including GJB2, AIFM1, and STRC were identified. The current study indicates that there are regional differences in the distribution of G6PD gene variation in AN, predominantly in the AN population of Guangxi. The potential mechanisms of hearing loss may be related to neonatal hyperbilirubinemia, genetic factors and others.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an enzymopathy that affects approximately 500 million people worldwide. A great number of mutations in the G6PD gene have been described. However, three class A G6PD variants known as G6PD Tomah (C385R), G6PD Kangnam (C385G), and G6PD Madrid (C385W) have been reported to be clinically important due to their associations with severe clinical manifestations such as hemolytic anemia. Therefore, this work aimed to perform, for the first time, biochemical and functional characterizations of these variants. The G6PD variants were cloned and purified for this purpose, followed by analyses of their kinetic parameters and thermal stability, as well as in silico studies. The results showed that the mutations induced changes in the proteins. Regarding the kinetic parameters, it was observed that the three variants showed lower affinities for G6P and NADP+, as well as lower thermal stability compared to WT-G6PD. Molecular dynamics simulations showed that C385 mutations induced changes around neighboring amino acids. Metadynamics simulations showed that most remarkable changes account for the binding pocket volumes, particularly in the structural NADP+ binding site, with a concomitant loss of affinity for catalytic processes.

Routine Screening of African American Males in Hematology Oncology Clinic for G6PD Deficiency: A Prospective Comparative Study of a Pharmacogenomic Panel Versus Standard Enzyme Assay

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common X-linked hereditary disorder in southern China. However, the incidence rate of G6PD deficiency and the frequency of the most common G6PD gene variants vary widely. The purpose of this study was to investigate the prevalence, genotype, and phenotypic features of G6PD deficiency in neonates in Fujian province, southeastern China. This retrospective cohort study enrolled 2,789,002 newborns (1,521,431 males and 1,267,571 females) based on the newborn screening program for G6PD deficiency in Fujian Province between January 2010 and December 2021. Of the 2,789,002 newborns enrolled, 26,437 cases were diagnosed (22,939 males and 3,498 females), and the estimated prevalence of G6PD deficiency in Fujian province was 0.95%. The prevalence was significantly higher among males (1.51%) than in females (0.28%) (p < 0.00001). Among the 3,198 patients with G6PD deficiency, 3,092 cases (2,145 males and 947 females) were detected to have G6PD gene variants. The top six prevalent genotypes identified represented 90.84% (2095/3,198) of the total and included c.1376G > T (44.93%), c.1388G > A (18.42%), c.1024C > T (9.32%), c.95A > G (8.69%), c.392G > T (5.25%), and c.871G > A (4.22%). The frequency of genotypes with c.1388G > A, c.1024C > T, and c.871G > A was higher in males in the Fujian province than in females, while the frequency of genotypes with c.1376G > T was lower. Furthermore, when comparing the enzyme activities of the top six prevalent genotypes, there were significant differences in the enzyme activities among the genotypes of male hemizygotes and female heterozygotes. According to the new classification of G6PD variants proposed by the World Health Organization (WHO), the variants with c.1376G > T, c.95A > G, and c.871G > A were recognized as Class A, while the c.392G > T, c.1388G > A, and c.1024C > T were recognized as Class B. To the best of our knowledge, this study is the first to systematically describe the overview of epidemiological characteristics of newborn G6PD deficiency in Fujian province, China, including the screening rate, incidence rate, and variant spectrum. Additionally, we elucidated the relationship between the distribution of enzyme activity with specific mutations and their WHO classification patterns. Our results could provide strategies for screening, diagnosis, and genetic counseling of G6PD deficiency in this area.

To investigate the prevalence, genotype and haematological characteristics of glucose-6-phosphate dehydrogenase (G6PD) deficiency in the blood donor population of Wuxi area (Jiangsu Province, China) and to assess the impact of their red blood cell (RBC) units on clinical transfusion. We conducted genotyping and large-scale screening for G6PD enzyme activity in the blood donors of Wuxi City. In addition, we assessed the haematological parameters of G6PD-deficient and non-deficient blood donors, and investigated the adverse transfusion reactions in patients transfused with G6PD-deficient blood. We investigated 17,113 blood donors, among whom 44 (0.26%) were tested positive for G6PD deficiency. We identified 40 G6PD gene variants, among which c.1388G>A, c.1376G>T, c.1024C>T and c.95A>G were common. In addition, we identified two novel G6PD gene variants, c.1312G>A and c.1316G>A. The G6PD-deficient and non-deficient blood samples showed a significant difference in the RBC, mean corpuscular volume (MCV), mean corpuscular Hb (MCH), RBC distribution width, total bilirubin (TBIL), direct bilirubin (DBIL) and indirect bilirubin (IBIL) values. However, the two samples showed no significant difference in the haemolysis rate at the end of the storage period. Finally, transfusion with G6PD-deficient RBC units did not lead to any adverse transfusion reactions. The positive rate of G6PD deficiency in the blood donor population of Wuxi City is 0.26%, and the genetic variants identified in this population are consistent with the common genetic variants observed in the Chinese population. Blood centres can establish a database on G6PD-deficient blood donors and mark their RBC units to avoid their use for special clinical patients.

Abstract Mutations in the X-chromosome G6PD gene cause glucose-6-phosphate dehydrogenase (G6PD) deficiency. Crohn’s disease (CD) is an inflammatory bowel disease that can cause inflammation and damage to the lining of the digestive tract. We report a 50-year-old male with CD who was found to have an unrecognized variant of G6PD deficiency. Further investigation confirmed the presence of G6PD enzymatic activity impairment, indicating the clinical relevance of this genetic alteration. This case report presents the G6PD variants rs137852339, rs1050757, and rs2230037 in an individual from South India, highlighting the significance of early genetic screening in individuals experiencing persistent symptoms lasting for years. The present case underscores the importance of genetic screening in patients with prolonged symptoms, aiming to prevent delayed diagnosis and enhance overall patient well-being through timely interventions and personalized treatment approaches.

The utilization of dried blood spot (DBS) samples for screening glucose-6-phosphate dehydrogenase (G6PD) deficiency remains limited in resource-constrained and malaria-endemic regions of Thailand. This study evaluated the feasibility of employing DBS for G6PD deficiency screening among 242 participants (118 males and 124 females) who provided fresh whole blood (FWB) and DBS samples for analysis. The G6PD deficiency diagnostic performances of the fluorescent spot test using DBS (FST-DBS) were compared to those using FWB (FST-FWB). The G6PD gene variant was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using DBS samples. The FST-DBS showed no significant difference in sensitivity (95.0% vs 100.0%) but superior specificity (100.0% vs 80.2%) and a positive predictive value (100.0% vs. 50.0%) compared to FST-FWB. The PCR-RFLP revealed a G6PD mutation incidence rate of 16.5% (11.1% in males and 5.4% in females). Among 40 DBS samples, 39 (97.5%) were the Mahidol (487G&gt;A) while a sample (2.5%) which both FST-DBS and FST-FWB showed positive results had an unidentified variant. Therefore, FST-DBS is an alternative format for G6PD deficiency screening in malaria-endemic regions, particularly where resources are limited.

Identification of glucose-6-phosphate dehydrogenase variants by utilizing polymerase chain reaction – Restriction fragment length polymorphism based method

Hyperbilirubinemia is a common disorder in neonates, with premature infants at higher risk of developing the disorder. Glucose-6-phosphate dehydrogenase (G6PD) gene detection was used to determine the incidence of G6PD deficiency and analyze the etiologies of G6PD deficiency in neonates with hyperbilirubinemia in the Zunyi region with the aim of providing scientific evidence for the clinical diagnosis and treatment. For the gene detection, 64 neonates with hyperbilirubinemia were selected as the observation group and 30 normal neonates were selected as the control group, and the risk factors for hyperbilirubinemia were investigated by using multivariate logistic regression analysis. Among the neonates in the observation group, 59 cases had the G1388A mutation (92.19%) and 5 cases had the G1376T mutation (7.81%). No mutation was detected in the control group. In the observation group, the proportion of neonates who were born prematurely, with artificial feeding, with the age of starting feeding of more than 24 h, the time of first bowel movement of more than 24 h, premature rupture of membranes, infection, scalp hematoma, and perinatal asphyxia was higher than that in the control group, and the difference was statistically significant (p< 0.05). Multivariate logistic regression analysis showed that prematurity, infection, scalp hematoma, perinatal asphyxia, the age of starting feeding of more than 24 h, and the time of first bowel movement over 24 h were risk factors for the development of neonatal hyperbilirubinemia (p< 0.05). The G1338A and G1376T mutations were important features of the genetics of neonatal hyperbilirubinemia, and genetic detection together with the prevention of prematurity, infection, scalp hematoma, perinatal asphyxia, the age of starting feeding, and the time of first bowel movement would help reduce the incidence of this disease.

Hanseniaspora uvarum is the predominant yeast species in the majority of wine fermentations, which has only recently become amenable to directed genetic manipulation. The genetics and metabolism of H. uvarum have been poorly studied as compared to other yeasts of biotechnological importance. This work describes the construction and characterization of homozygous deletion mutants in the HuZWF1 gene, encoding glucose-6-phosphate dehydrogenase (G6PDH), which provides the entrance into the oxidative part of the pentose phosphate pathway (PPP) and serves as a major source of NADPH for anabolic reactions and oxidative stress response. Huzwf1 deletion mutants grow more slowly on glucose medium than wild-type and are hypersensitive both to hydrogen peroxide and potassium bisulfite, indicating that G6PDH activity is required to cope with these stresses. The mutant also requires methionine for growth. Enzyme activity can be restored by the expression of heterologous G6PDH genes from other yeasts and humans under the control of a strong endogenous promoter. These findings provide the basis for a better adaptation of H. uvarum to conditions used in wine fermentations, as well as its use for other biotechnological purposes and as an expression organism for studying G6PDH functions in patients with hemolytic anemia.

Introduction: Patients with acute myeloid leukemia (AML) show a higher risk for several types of infections, including fungal infections (FI), which are one of the main causes of morbidity and mortality. Glucose-6-phosphate dehydrogenase (G6PD) is an enzyme located in all cells that is very necessary in leukocytes for the production of basic and acid proteases that are used to destroy invading microorganisms. Our objective in this study was to evaluate whether polymorphisms in the G6PD gene concomitantly with FI are associated with clinical events and morbidity in patients diagnosed with AML and followed up at the Amazonas State Blood Center (HEMOAM), Manaus, Brazil. Materials and Methods: The study population was randomly constituted of adults and children, of either sex, and any age, with a diagnosis of acute myeloid leukemia, all of whom were undergoing treatment at the HEMOAM. Molecular genotyping was performed using real-time PCR (qPCR) and subsequent Sanger sequencing to confirm the c.202G &gt; A/c.376A &gt; G polymorphisms. Results: A total of 157 patients (91 (58%) males and 66 (42%) females) were involved in the study. The most prevalent AML subtype in the studied group was M3 in 63 patients (40.12%), followed by M5 in 33 patients (21.02%), M2 in 21 patients (13.37%) and M4 in 15 patients (9.55%), with a similar prevalence between genders. The prevalence of fungal infections was identical between genders; however, bruising (p = 0.004), vomiting (p = 0.016) and cardiac alterations (p &lt; 0.001) were higher in females, while persistent cough (p = 0.049) and diarrhea (p &lt; 0.001) were higher in males. A total of eighteen patients presents G6PD polymorphisms, with 8 (5.1%) of these for c.202GA/AA, 18 (11.5%) for c.376AG/GG and 4 (2.5%) for both polymorphisms concomitantly (c.202AA/c.376GG). However, the prevalence of death in patients affected with FI was much higher in those that have these polymorphisms (p &lt; 0.001). Conclusion: We believe that the determination of G6PD polymorphisms will allow the development of monitoring strategies, and aid in early diagnosis and the appropriate and targeted treatment for AML. In addition, evaluating their activity may help to identify AML patients at a higher risk of FI, thus allowing the design of more intensive therapeutic and surveillance strategies.