• Title/Summary/Keyword: rGH

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Highly Efficient Biotransformation of Notoginsenoside R1 into Ginsenoside Rg1 by Dictyoglomus thermophilum β-xylosidase Xln-DT

  • Li, Qi;Wang, Lei;Fang, Xianying;Zhao, Linguo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.4
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    • pp.447-457
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    • 2022
  • Notoginsenoside R1 and ginsenoside Rg1 are the main active ingredients of Panax notoginseng, exhibiting anti-fatigue, anti-tumor, anti-inflammatory, and other activities. In a previous study, a GH39 β-xylosidase Xln-DT was responsible for the bioconversion of saponin, a natural active substance with a xylose group, with high selectivity for cleaving the outer xylose moiety of notoginsenoside R1 at the C-6 position, producing ginsenoside Rg1 with potent anti-fatigue activity. The optimal bioconversion temperature, pH, and enzyme dosage were obtained by optimizing the transformation conditions. Under optimal conditions (pH 6.0, 75℃, enzyme dosage 1.0 U/ml), 1.0 g/l of notoginsenoside R1 was converted into 0.86 g/l of ginsenoside Rg1 within 30 min, with a molar conversion rate of approximately 100%. Furthermore, the in vivo anti-fatigue activity of notoginsenoside R1 and ginsenoside Rg1 were compared using a suitable rat model. Compared with the control group, the forced swimming time to exhaustion was prolonged in mice by 17.3% in the Rg1 high group (20 mg/kg·d). Additionally, the levels of hepatic glycogen (69.9-83.3% increase) and muscle glycogen (36.9-93.6% increase) were increased. In the Rg1 group, hemoglobin levels were also distinctly increased by treatment concentrations. Our findings indicate that treatment with ginsenoside Rg1 enhances the anti-fatigue effects. In this study, we reveal a GH39 β-xylosidase displaying excellent hydrolytic activity to produce ginsenoside Rg1 in the pharmaceutical and food industries.

Factors Affecting on Final Adult Height and Total Height Gain in Children with Idiopathic and Organic Growth Hormone Deficiency after Growth Hormone Treatment (특발성과 기질성 성장호르몬 결핍증 환아에서 성장호르몬 치료 후 최종 성인신장과 신장 증가에 영향을 미치는 인자)

  • Choi, Im Jeong;Hwang, Jin Soon;Shin, Choong Ho;Yang, Sei Won
    • Clinical and Experimental Pediatrics
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    • v.46 no.8
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    • pp.803-810
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    • 2003
  • Purpose : The purpose of this study was to evaluate the factors affecting the final adult height and total height gain in idiopathic and organic growth hormone deficient(GHD) children after growth hormone(GH) treatment. Methods : Thirteen patients with idiopathic GHD and 22 patients with organic GHD who had been treated with GH and attained adult final height were included in this study. Factors which could affect the final adult height(FAH) and total height gain, were evaluated. Results : Height SDS(standard deviation score) at initial GH treatment in idiopathic GHD was significantly shorter than that in organic GHD($-4.13{\pm}1.28$ vs $-1.66{\pm}1.06$, P<0.001). Growth velocity during the first year of GH treatment was $9.69{\pm}3.19cm$(idiopathic GHD) and $7.87{\pm}3.65cm$(organic GHD). Height(SDS) at puberty in organic GHD was significantly greater than in idiopathic GHD ($-0.55{\pm}1.25$ vs $-2.28{\pm}0.95$, P<0.001). Final adult height(SDS) was significantly greater in organic GHD than in idiopathic GHD($0.22{\pm}1.06$ vs $-1.44{\pm}0.84$, P<0.001). In idiopathic GHD, total height gain (SDS) was most significantly correlated with midparental height minus initial height(MPH-IH)(SDS) (r=0.886, P<0.001). Total height gain(SDS) was more significantly correlated with MPH-IH(SDS) and prepubertal height gain(SDS) in idiopathic GHD(r=0.640, P=0.01, r=0.801, P<0.001). Conclusion : Final adult height was greater in organic GHD than in idiopathic GHD patients. While total height gain(SDS) was more pronounced in children with lower initial height compared to MPH, absolute final adult height was influenced by height at puberty. To improve the final adult height in children with GHD, height at onset of puberty must be increased by early diagnosis and continuous treatment with optimal doses of GH. There results should be evaluated with more patients.

Gene Cloning, Expression, and Characterization of a $\beta$-Agarase, AgaB34, from Agarivorans albus YKW-34

  • Fu, Xiao Ting;Pan, Cheol-Ho;Lin, Hong;Kim, Sang-Moo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.257-264
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    • 2009
  • A $\beta$-agarase gene, agaB34, was functionally cloned from the genomic DNA of a marine bacterium, Agarivorans albus YKW-34. The open reading frame of agaB34 consisted of 1,362 bp encoding 453 amino acids. The deduced amino acid sequence, consisting of a typical N-terminal signal peptide followed by a catalytic domain of glycoside hydrolase family 16 (GH-16) and a carbohydrate-binding module (CBM), showed 37-86% identity to those of agarases belonging to family GH-16. The recombinant enzyme (rAgaB34) with a molecular mass of 49 kDa was produced extracellularly using Escherichia coli $DH5{\alpha}$ as a host. The purified rAgaB34 was a $\beta$-agarase yielding neoagarotetraose (NA4) as the main product. It acted on neoagarohexaose to produce NA4 and neoagarobiose, but it could not further degrade NA4. The maximal activity of rAgaB34 was observed at $30^{\circ}C$ and pH 7.0. It was stable over pH 5.0-9.0 and at temperatures up to $50^{\circ}C$. Its specific activity and $k_{cat}/K_m$ value for agarose were 242 U/mg and $1.7{\times}10^6/sM$, respectively. The activity of rAgaB34 was not affected by metal ions commonly existing in seawater. It was resistant to chelating reagents (EDTA, EGTA), reducing reagents (DTT, $\beta$-mercaptoethanol), and denaturing reagents (SDS and urea). The E. coli cell harboring the pUC18-derived agarase expression vector was able to efficiently excrete agarase into the culture medium. Hence, this expression system might be used to express secretory proteins.

Molecular Cloning, Overexpression, and Enzymatic Characterization of Glycosyl Hydrolase Family 16 ${\beta}$-Agarase from Marine Bacterium Saccharophagus sp. AG21 in Escherichia coli

  • Lee, Youngdeuk;Oh, Chulhong;Zoysa, Mahanama De;Kim, Hyowon;Wickramaarachchi, Wickramaarachchige Don Niroshana;Whang, Ilson;Kang, Do-Hyung;Lee, Jehee
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.913-922
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    • 2013
  • An agar-degrading bacterium was isolated from red seaweed (Gelidium amansii) on a natural seawater agar plate, and identified as Saccharophagus sp. AG21. The ${\beta}$-agarase gene from Saccharophagus sp. AG21 (agy1) was screened by long and accurate (LA)-PCR. The predicted sequence has a 1,908 bp open reading frame encoding 636 amino acids (aa), and includes a glycosyl hydrolase family 16 (GH16) ${\beta}$-agarase module and two carbohydrate binding modules of family 6 (CBM6). The deduced aa sequence showed 93.7% and 84.9% similarity to ${\beta}$-agarase of Saccharophagus degradans and Microbulbifer agarilyticus, respectively. The mature agy1 was cloned and overexpressed as a His-tagged recombinant ${\beta}$-agarase (rAgy1) in Escherichia coli, and had a predicted molecular mass of 69 kDa and an isoelectric point of 4.5. rAgy1 showed optimum activity at $55^{\circ}C$ and pH 7.6, and had a specific activity of 85 U/mg. The rAgy1 activity was enhanced by $FeSO_4$ (40%), KCl (34%), and NaCl (34%), compared with the control. The newly identified rAgy1 is a ${\beta}$-agarase, which acts to degrade agarose to neoagarotetraose (NA4) and neoagarohexaose (NA6) and may be useful for applications in the cosmetics, food, bioethanol, and reagent industries.

Heterogeneity in liver histopathology is associated with GSK-3β activity and mitochondrial dysfunction in end-stage diabetic rats on differential diets

  • Lee, Jun-Ho;Choi, Soo-Bong;Sung, Dong-Jun;Jin, Mingli;Lee, Ju-Han;Mun, Ji-Young;Hwang, Tae-Sook;Han, Sang-Don;Ro, Young-Tae;Kim, Sung-Young;You, Jueng-Soo;Lim, Inja;Noh, Yun-Hee
    • BMB Reports
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    • v.53 no.2
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    • pp.100-105
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    • 2020
  • While liver histopathology is heterogeneous in diabetes, the underlying mechanisms remain unclear. We investigated whether glycemic variation resulting from differential diets can induce heterogeneity in diabetic liver and the underlying molecular mechanisms. We generated end-stage non-obese diabetic model rats by subtotal-pancreatectomy in male Sprague-Dawley rats and ad libitum diet for 7 weeks (n = 33). The rats were then divided into three groups, and fed a standard- or a low-protein diet (18 or 6 kcal%, respectively), for another 7 weeks: to maintain hyperglycemia, 11 rats were fed ad libitum (18AL group); to achieve euglycemia, 11 were calorie-restricted (18R group), and 11 were both calorie- and protein-restricted with the low-protein diet (6R group). Overnight-fasted liver samples were collected after the differential diets together with sham-control (18S group), and histology and molecular changes were compared. Hyperglycemic-18AL showed glycogenic hepatopathy (GH) without steatosis, with the highest GSK-3β inactivation because of Akt activation during hyperglycemia; mitochondrial function was not impaired, compared to the 18S group. Euglycemic-18R showed neither GH nor steatosis, with intermediate GSK-3β activation and mitochondrial dysfunction. However, euglycemic-6R showed both GH and steatosis despite the highest GSK-3β activity and no molecular evidence of increased lipogenesis or decreased ApoB expression, where mitochondrial dysfunction was highest among the groups. In conclusion, heterogeneous liver histopathology developed in end-stage non-obese diabetic rats as the glycemic levels varied with differential diets, in which protein content in the diets as well as glycemic levels differentially influenced GSK-3β activity and mitochondrial function in insulin-deficient state.

Bovine Growth Hormone and Milk Fat Synthesis: from the Body to the Molecule - Review -

  • Kim, W.Y.;Ha, J.K.;Han, In K.;Baldwin, R.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.4
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    • pp.335-356
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    • 1997
  • Injection of bovine growth hormone (bGH) to lactating dairy cows increases milk yield and yields of milk components including fat. It is generally believed that most of the anabolic effects derived from bGH in animal tissues are primarily mediated by IGF-1. IGF-1 is a strong anabolic peptide in the plasma of animals and exerts mitogenic and metabolic effects on target cells. Contrary to most protein hormones, the majority of IGF-1 in circulation is bound to the binding proteins (IGFBPs) which are known to be responsible for modifying the biological actions of IGF-1, thus making determinations of IGF-1 actions more difficult. On the other hand, fat is a major milk component and the greatest energy source in milk. Currently, the fat content of milk is one of the major criteria used in determining milk prices. It has been known that flavor and texture of dairy products are mainly affected by milk fat and its composition. Acetyl-CoA carboxylase (ACC) is the rate limiting enzyme which catalyzes the conversion of acetyl-CoA to malonyl-CoA for fatty acid synthesis in 1ipogenic tissues of animals including bovine lactating mammary glands. In addition to the short-tenn hormonal regulation of ACC by changes in the catalytic efficiency per enzyme molecule brought about by phosphorylation and dephosphorylation of the enzyme, the long-term hormonal regulation of ACC by changes in the number of enzyme molecules plays an essential role in control of ACC and lipogenesis. Insulin, at supraphysiological concentrations, binds to IGF-1 receptors, thereby mimicking the biological effects of IGF-1. The receptors for insulin and IGF-1 share structural and functional homology. Furthermore, epidermal growth factor increased ACC activity in rat hepatocytes and adipocytes. Therefore, it can be assumed that IGF-1 mediating bGH action may increase milk fat production by stimulation ACC with phosphorylation (short term) and/or increasing amounts of the enzyme proteins (long term). Consequently, the main purpose of this paper is to give the readers not only the galactopoietic effects of bGH, but also the insight of bGH action with regard to stimulating milk fat synthesis from the whole body to the molecular levels.

Effects of Intra-duodenal Infusion of Limiting Amino Acids on Plasma Insulin-like Growth Factor I, Growth Hormone, Insulin and Liver Insulin-like Growth Factor I mRNA in Growing Goat Fed a Maize Stover-based Diet

  • Sun, Z.H.;Tan, Z.L.;Yao, J.H.;Tang, Z.R.;Shan, J.G.;Hu, J.P.;Tang, S.X.;Jiang, Y.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.733-741
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    • 2007
  • The effects of intra-duodenal infusion of methionine (Met), lysine (Lys) and leucine (Leu) on dry matter intake (DMI), the concentrations of insulin-like growth factor I (IGF-I), growth hormone (GH) and insulin in plasma, and liver IGF-I mRNA level were investigated in two experiments for Liuyang Black growing wether goats. In Experiment 1, three goats ($10.0{\pm}0.1$ kg) were fitted with ruminal, proximal duodenal and terminal ileal fistulaes to determine the infusion amounts of Met, Lys and Leu at the duodenum according to essential amino acid flows into the duodenum and their apparent digestibility. The infusion amounts were 0.77 g/d, 0.91 g/d and 0.58 g/d respectively. In Experiment 2, 4 groups of goats (($10.0{\pm}0.2$ kg) for each group, were cannulated at the duodenum, and were infused with a mixture of Met, Lys and Leu (Control), or mixtures with 21% Met, Lys or Leu replaced with glutamate respectively on a nitrogenous basis. The replacement of 21% Met, Lys or Leu with glutamate did not affect intakes of maize stover, concentrate or both (p>0.05) when compared with the control. The replacement of 21% Met or Lys significantly (p<0.05) reduced plasma GH, insulin and IGF-I concentrations and liver IGF-I mRNA level. The replacement of 21% Leu with glutamate reduced (p<0.05) plasma IGF-I concentration only, but not plasma insulin and GH, as well as liver IGF-I mRNA level (p>0.05). The close relationships between supplying Met and Lys in the lumen of the duodenum and plasma IGF-I, GH and insulin concentrations, as well as liver IGF-I mRNA level in this study indicate that the effects of the limiting amino acids on nutrition of animals are likely intermediated via their effects on these hormones, and these hormone profiles could be used as intermediate markers for the limiting order of amino acids.

Analyzing environmental factors influencing the gestation length and birth weight of Hanwoo cattle

  • Shil Jin;Sung-Sik Kang;Jeong Il Won;Hyoun Ju Kim;Sun Sik Jang;Sung Woo Kim
    • Journal of Animal Reproduction and Biotechnology
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    • v.39 no.2
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    • pp.121-130
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    • 2024
  • Background: This study focused on reproductive traits in Hanwoo cattle, specifically the environmental factors affecting gestation length and birth weight. Methods: The records of 1,540 cows calved at the Hanwoo Research Institute from 2015 to 2023 were examined. This study analyzed two populations, line-breeding Hanwoo (LBH) and general Hanwoo (GH), with all cows undergoing estrus synchronization and artificial insemination. The R software was used to compare the differences between the two populations and analyze the environmental factors affecting each trait. Results: The results showed that the average gestation length for LBH was 283.28 ± 5.93 days, which was significantly shorter than that of the GH, which had an average of 285.63 ± 6.21 days (p < 0.001). The average birth weight of LBH calves was 25.10 ± 3.69 kg, significantly lighter than GH calves, which weighed 27.26 ± 4.11 kg on average (p < 0.001). Analysis of environmental factors revealed significant differences in the gestation length of LBH based on dam parity, year, and season of calving. However, no significant differences were observed based on calf sex. For LBH, birth weight showed significant differences based on dam parity, year of calving, and sex of the calf, but not the season of calving. In GH, gestation length varied with dam parity and calving season, but not with calving year or calf sex. The GH birth weight showed differences based on dam parity, year of calving, and calf sex, but not the season of calving. Conclusions: Reproductive traits in the Hanwoo cattle industry are economically vital but are heavily influenced by environmental factors due to their low heritability. An accurate evaluation of the genetic potential of these traits requires an analysis of the environmental factors affecting them. The results of this study serve as foundational data for predicting the potential for genetic improvement in the gestation length and birth weight of Hanwoo cattle.

Mechanical Characteristics of the Rift, Grain and Hardway Planes in Jurassic Granites, Korea (쥬라기 화강암류에서 발달된 1번 면, 2번 면 및 3번 면의 역학적 특성)

  • Park, Deok-Won
    • Korean Journal of Mineralogy and Petrology
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    • v.33 no.3
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    • pp.273-291
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    • 2020
  • The strength characteristics of the three orthogonal splitting planes, known as rift, grain and hardway planes in granite quarries, were examined. R, G and H specimens were obtained from the block samples of Jurassic granites in Geochang and Hapcheon areas. The directions of the long axes of these three specimens are perpendicular to each of the three planes. First, The chart, showing the scaling characteristics of three graphs related to the uniaxial compressive strengths of R, G and H specimens, were made. The graphs for the three specimens, along with the increase of strength, are arranged in the order of H < G < R. The angles of inclination of the graphs for the three specimens, suggesting the degree of uniformity of the texture within the specimen, were compared. The above angles for H specimens(θH, 24.0°~37.3°) are the lowest among the three specimens. Second, the scaling characteristics related to the three graphs of RG, GH and RH specimens, representing a combination of the mean compressive strengths of the two specimens, were derived. These three graphs, taking the various N-shaped forms, are arranged in the order of GH < RH < RG. Third, the correlation chart between the strength difference(Δσt) and the angle of inclination(θ) was made. The above two parameters show the correlation of the exponential function with an exponent(λ) of -0.003. In both granites, the angle of inclination(θRH) of the RH-graph is the lowest. Fourth, the six types of charts, showing the correlations among the three kinds of compressive strengths for the three specimens and the five parameters for the two sets of microcracks aligned parallel to the compressive load applied to each specimen, were made. From these charts for Geochang and Hapcheon granites, the mean value(0.877) of the correlation coefficients(R2) for total density(Lt), along with the frequency(N, 0.872) and density(ρ, 0.874), is the highest. In addition, the mean values(0.829) of correlation coefficients associated with the mean compressive strengths are more higher than the minimum(0.768) and maximum(0.804) compression strengths of three specimens. Fifth, the distributional characteristics of the Brazilian tensile strengths measured in directions parallel to the above two sets of microcracks in the three specimens from Geochang granite were derived. From the related chart, the three graphs for these tensile strengths corresponding to the R, G and H specimens show an order of H(R1+G1) < G(R2+H1) < R(R1+G1). The order of arrangement of the three graphs for the tensile strengths and that for the compressive strengths are mutually consistent. Therefore, the compressive strengths of the three specimens are proportional to the three types of tensile strengths. Sixth, the values of correlation coefficients, among the three tensile strengths corresponding to each cumulative number(N=1~10) from the above three graphs and the five parameters corresponding to each graph, were derived. The mean values of correlation coefficients for each parameter from the 10 correlation charts increase in the order of density(0.763) < total length(0.817) < frequency(0.839) < mean length(Lm, 0.901) ≤ median length(Lmed, 0.903). Seventh, the correlation charts among the compressive strengths and tensile strengths for the three specimens were made. The above correlation charts were divided into nine types based on the three kinds of compressive strengths and the five groups(A~E) of tensile strengths. From the related charts, as the tensile strength increases with the mean and maximum compressive strengths excluding the minimum compressive strength, the value of correlation coefficient increases rapidly.

Characterization of $\beta$-Ketoadipate Pathway from Multi-Drug Resistance Bacterium, Acinetobacter baumannii DU202 by Proteomic Approach

  • Park, Soon-Ho;Kim, Jae-Woo;Yun, Sung-Ho;Leem, Sun-Hee;Kahng, Hyung-Yeel;Kim, Seung-Il
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.632-640
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    • 2006
  • In this study, the biodegradative activities of monocyclic aromatic compounds were determined from the multi-drug resistant (MDR) Acinetobacter baumannii, which were studied in the form of clinical isolates from a hospital in Korea. These bacteria were capable of biodegrading monocyclic aromatic compounds, such as benzoate and p-hydroxybenzoate. In order to determine which pathways are available for biodegradation in these stains, we conducted proteome analyses of benzoate, and p-hydroxybenzoate-cultured A. baumannii DU202, using 2-DE/MS analysis. As genome DB of A. baumannii was not yet available, MS/MS analysis or de novo sequencing methods were employed in the identification of induced proteins. Benzoate branch enzymes [catechol 1,2-dioxygenase (CatA) and benzoate dioxygenase $\alpha$ subunit (BenA)] of the $\beta$-ketoadipate pathway were identified under benzoate culture condition and p-hydroxybenzoate branch enzymes [protocatechuate 3,4-dioxygenas $\alpha$ subunit (PcaG) and 3-carboxy-cis,cis-muconate cycloisomerase (PcaR)] of the $\beta$-ketoadipate pathway were identified under p-hydroxybenzoate culture condition, respectively, thereby suggesting that strain DU202 utilized the $\beta$-ketoadipate pathway for the biodegradation of monocyclic aromatic compounds. The sequence analysis of two purified dioxygenases (CatA and PcaGH) indicated that CatA is closely associated with the CatA of Acinetobacter radiresistance, but PcaGH is only moderately associated with the PcaGH of Acinetobacter sp. ADPI. Interestingly, the fused form of PcaD and PcaC, carboxymuconolactone decarboxylase (PcaCD), was detected on benzoate-cultured A. baumannii DU202. These results indicate that A. baumannii DU202 exploits a different $\beta$-ketoadipate pathway from other Acinetobacter species.