Cloning and nucleotide sequence of the structural genes encoding the formate dehydrogenase of Wolinella succinogenes

The formate dehydrogenase of Wolinella succinogenes is a membraneous molybdo-enzyme which is involved in phosphorylative electron transport. The gene (fdhA) encoding the largest subunit was isolated from a gene bank by immunological screening. The fdhA gene was located in an apparent transcriptional unit (fdh A, B, C. D) together with three more structural genes. The N-terminal sequences of three polypeptides present in the isolated enzyme were found to map within the fdhA, B and C structural genes. A polypeptide corresponding to fdhD was not detected in the enzyme preparation. This suggested that the functional formate dehydrogenase was made up of three or four different subunits. The genes fdhA and C encode larger preproteins which differ from the corresponding mature proteins by N-terminal signal peptides. The N-terminal half of the mature FdhA is homologous to the larger subunits of the formate dehydrogenases of E. coli (formate-hydrogenlyase linked) and Methanobacterium formicicum as well as to three bacterial reductases containing molybdenum. It harbours a conserved cysteine cluster and two more domains which may be involved in binding the molybdenum cofactor. FdhB may represent an iron-sulphur protein, twelve cysteine residues of which are arranged in two clusters which are typical of ligands of the iron-sulfur centers in ferredoxins. FdhC is a hydrophobic protein with four predicted transmembrane segments, which appears to be identical with the cytochrome b present in the isolated formate dehydrogenase. It may form the membrane anchor of the enzyme and react with the bacterial menaquinone

Cloning and nucleotide sequence of the psrA gene of Wolinella succinogenes polysulphide reductase

The polysulphide reductase (formerly sulphur reductase) of Wolinella succinogenes is a component of the phosphorylative electron transport system with polysulphide as the terminal acceptor. Using an antiserum raised against the major subunit (PsrA, 85 kDa) of the enzyme, the corresponding gene (psrA) was cloned from a lambda-gene bank. The N-terminal amino acid sequence of PsrA mapped within the psrA gene product, which also contained an apparent signal peptide. Downstream of the psrA gene two more open reading frames (psrB and psrC) were found. The three genes may form a transcriptional unit with the transcription start site in front of psrA. The three genes were present only once on the genome. PsrA is a hydrophilic protein homologous to the largest subunits of six prokaryotic molybdoenzymes. PsrB is predicted to be hydrophilic, to contain ferredoxin-like cysteine clusters and to be homologous to the smaller hydrophilic subunits of four molybdoenzymes. PsrC is predicted to be a hydrophobic protein that could possibly serve as the membrane anchor of the enzyme

Myoelectric or Force Control? A Comparative Study on a Soft Arm Exosuit

The intention-detection strategy used to drive an exosuit is fundamental to evaluate the effectiveness and acceptability of the device. Yet, current literature on wearable soft robotics lacks evidence on the comparative performance of different control approaches for online intention-detection. In the present work, we compare two different and complementary controllers on a wearable robotic suit, previously formulated and tested by our group; a model-based myoelectric control (myoprocessor), which estimates the joint torque from the activation of target muscles, and a force control that estimates human torques using an inverse dynamics model (dynamic arm). We test them on a cohort of healthy participants performing tasks replicating functional activities of daily living involving a wide range of dynamic movements. Our results suggest that both controllers are robust and effective in detecting human–motor interaction, and show comparable performance for augmenting muscular activity. In particular, the biceps brachii activity was reduced by up to 74% under the assistance of the dynamic arm and up to 47% under the myoprocessor, compared to a no-suit condition. However, the myoprocessor outperformed the dynamic arm in promptness and assistance during movements that involve high dynamics. The exosuit work normalized with respect to the overall work was $68.84 pm 3.81$ when it was ran by the myoprocessor, compared to $45.29 pm 7.71$ during the dynamic arm condition. The reliability and accuracy of motor intention detection strategies in wearable device is paramount for both the efficacy and acceptability of this technology. In this article, we offer a detailed analysis of the two most widely used control approaches, trying to highlight their intrinsic structural differences and to discuss their different and complementary performance

Rigid, Soft, Passive, and Active: A Hybrid Occupational Exoskeleton for Bimanual Multijoint Assistance

Physically demanding work is still common in western countries, with large proportions of the workforce that are exposed for more than a quarter of their working time to tiring postures or repetitive tasks: the shoulder is one of the main body areas susceptible to work-related musculo-skeletal disorders. Recent advancements in assistive technology have provided new instruments to promote safety and reduce workload. Colloquially referred to as occupational exoskeletons (OEs), these wearable devices are usually spring-loaded, and provide gravity support for overhead tasks. OEs for upper limbs are usually single-joint exoskeletons and assist shoulder flexion/extension; they do not provide support to distal joints such as the elbow. In the present work, starting from a commercially available exoskeleton, we propose an innovative concept of hybrid upper-limb OEs. We combined a spring-loaded shoulder exoskeleton with an active elbow exosuit to extend the capability of the OEs to provide gravitational support to both shoulder and elbow flexion-extension in strenuous manual tasks. The proposed device can reduce up to 32% of the biceps activity during the elbow flexion and up to 31% of the deltoids activity during the shoulder abduction. In-lab experimentation showed the potentials of such a hybrid approach in reducing the strain of the upper-limb muscles