High-throughput process development for biopharmaceutical drug substances

Quality by Design (QbD) is gaining industry acceptance as an approach towards development and commercialization of biotechnology therapeutic products that are expressed via microbial or mammalian cell lines. In QbD, the process is designed and controlled to deliver specified quality attributes consistently. To acquire the enhanced understanding that is necessary to achieve the above, however, requires more extensive experimentation to establish the design space for the process and the product. With biotechnology companies operating under ever-increasing pressure towards lowering the cost of manufacturing, the use of high-throughput tools has emerged as a necessary enabler of QbD in a time- and resource-constrained environment. We review this topic for those in academia and industry that are engaged in drug substance process development.     

The role of biotechnology in art preservation

Biotechnology has played a key role in medicine, agriculture and industry for over 30 years and has advanced our understanding of the biological sciences. Furthermore, the tools of biotechnology have a great and largely untapped potential for the preservation and restoration of our cultural heritage. It is possible that these tools are not often applied in this context because of the inherent separation of the worlds of art and science; however, it is encouraging to see that during the past six years important biotechnological applications to artwork preservation have emerged and advances in biotechnology predict further innovation. In this article we describe and reflect upon a unique example of a group of scientists and art restoration technicians working together to study and treat of a piece of colonial art, and review some of the new applications in biotechnology for the preservation of mankind's cultural heritage. We predict an expansion in this field and the further development of biotechnological techniques, which will open up new opportunities to both biologists and artwork preservers.     

An image method to evaluate bagasse fiber dimensions

In the last 25 years the amount of textile nonwovens used for industrial and commercial applications increased more than 10 times. Bagasse fiber, a by-product from sugar cane industry, provides a natural resource for nonwoven industries. Even though underrated as a potential fiber, bagasse comes more and more into attention because of the increasing concern for disposal of agricultural residuals and the need for enhancing the sugar cane industry’s profitability. However, there is a lack of an instrumental method to evaluate bagasse fiber length and fineness. This paper presents a study on measuring the bagasse fineness using image analysis method. Cross-sections images of bagasse fibers were visualized using Scanning Electronic Microscopy (SEM). The computing software Scion Image was used to measure bagasse fiber length and cross-sectional area. Relationship between fiber fineness and cross-sectional area was analyzed using the statistical method of regression.     

Research and application of biotechnology in textile industries in China

Textile industry is a conventional and pillar industry in China, which possesses a considerable proportion of the national economy. In recent years, special attention has been paid to the application of biotechnology in textile industries in China. As an interdiscipline between natural science and engineering science, textile biotechnology has much effect on China's textile industry. This paper summarizes current developments and highlights those areas where biotechnology might play an increasingly important role in China's textile industry as follows:

(1) Development of new types of textile fibers and polymers, such as Bt cotton naturally colored cotton, colored silk and silk gene-sequence, spider silk non-wovens, chitin fiber and chitosan derivatives, etc.

(2) Application of enzyme technology in textile wet processing, such as alkaline pectinase, PVA-degrading enzyme, cutinase and catalase used for cotton preparation, neutral cellulase for denim washing, transglutaminase for wool modification, protease for silk degumming as well as pectinase and hemicellulases for retting of bast fibers.

(3) Treatment of textile effluents with biotechnology.

Next‐Generation Industrial Biotechnology‐Transforming the Current Industrial Biotechnology into Competitive Processes

The chemical industry has made a contribution to modern society by providing cost‐competitive products for our daily use. However, it now faces a serious challenge regarding environmental pollutions and greenhouse gas emission. With the rapid development of molecular biology, biochemistry, and synthetic biology, industrial biotechnology has evolved to become more efficient for production of chemicals and materials. However, in contrast to chemical industries, current industrial biotechnology (CIB) is still not competitive for production of chemicals, materials, and biofuels due to their low efficiency and complicated sterilization processes as well as high‐energy consumption. It must be further developed into “next‐generation industrial biotechnology” (NGIB), which is low‐cost mixed substrates based on less freshwater consumption, energy‐saving, and long‐lasting open continuous intelligent processing, overcoming the shortcomings of CIB and transforming the CIB into competitive processes. Contamination‐resistant microorganism as chassis is the key to a successful NGIB, which requires resistance to microbial or phage contaminations, and available tools and methods for metabolic or synthetic biology engineering. This review proposes a list of contamination‐resistant bacteria and takes Halomonas spp. as an example for the production of a variety of products, including polyhydroxyalkanoates under open‐ and continuous‐processing conditions proposed for NGIB.     

Liquid and gaseous fuels from biotechnology: challenge and opportunities

Abstract: This paper presents challenging opportunities for production of liquid and gaseous fuels by biotechnology. From the liquid fuels, ethyl alcohol production has been widely researched and implemented. The major obstacle for large scale production of ethanol for fuel is the cost, whereby the substrate represents one of the major cost components. Various scenarios will be presented for a critical assessment of cost distribution for production of ethanol from various substrates by conventional and high rate processes. The paper also focuses on recent advances in the research and application of biotechnological processes and methods for the production of liquid transportation fuels other than ethanol (other oxygenates; diesel fuel extenders and substitutes), as well as gaseous fuels (biogas, methane, reformed syngas). Potential uses of these biofuels are described, along with environmental concerns which accompany them. Emphasis is also put on microalgal lipids as diesel substitute and biogas/methane as a renewable alternative to natural gas. The capturing and use of landfill gases is also mentioned, as well as microbial coal liquefaction. Described is also the construction and performance of microbial fuel cells for the direct high-efficiency conversion of chemical fuel energy to electricity. Bacterial carbon dioxide recovery is briefly dealt with as an environmental issue associated with the use of fossil energy.     

Influence of the application of sugarcane bagasse on lindane (gamma-HCH) mobility through soil column: implication for biotreatment

In the present study we employed sugarcane bagasse for biotreatment of soil containing 50mgkg(-1) of lindane. Garden soil were treated with lindane and amended with varying concentrations of sugarcane bagasse (10%, 20%, 30%, 40% and 50%; w/w). Data on dissipation and degradation of lindane in soil columns (0-15, 15-30cm) were studied at six consecutive samplings (0, 3, 7, 45 and 60 days). Treatment with 50% sugarcane bagasse resulted in >53% degradation of lindane in upper soil column with minimal leaching to lower soil column (0.002%) while highest leaching of lindane from upper soil column to lower soil column was occurred in garden soil (35.8%). Similarly, a substantial microbial biomass input has detected in amended soil than garden soil. Our results provide evidence that sugarcane bagasse can accelerate lindane degradation by enhanced microbial activity and prevent pesticide mobility through soil column by adsorption. Sugarcane bagasse could be useful as cheaper, easy available alternative for the biotreatment of lindane impacted soil.     

Analysis of pH-induced population oscillations of Saccharomyces cerevisiae and Escherichia coli using photon correlation spectroscopy

Following a recent successful application of p.c.s. to liquid chromatography in the biotechnology industry, its usefulness as a contamination monitor in the fermentation industry was assessed. It was found that: (i) the intensity bias of the technique limits its uses to the detection of contaminants when they are larger than the host; (ii) the inherent heterogeneity of microbial cultures prevents the use of multiangle studies, and (iii) the large size of bacteria make the use of p.c.s. in flowing, on-line systems impractical.     

Use of molecular techniques in bioremediation

In a practical sense, biotechnology is concerned with the production of commercial products generated by biological processes. More formally, biotechnology may be defined as "the application of scientific and engineering principles to the processing of material by biological agents to provide goods and services" (Cantor, 2000). From a historical perspective, biotechnology dates back to the time when yeast was first used for beer or wine fermentation, and bacteria were used to make yogurt. In 1972, the birth of recombinant DNA technology moved biotechnology to new heights and led to the establishment of a new industry. Progress in biotechnology has been truly remarkable. Within four years of the discovery of recombinant DNA technology, genetically modified organisms (GMOs) were making human insulin, interferon, and human growth hormone. Now, recombinant DNA technology and its products--GMOs are widely used in environmental biotechnology (Glick and Pasternak, 1988; Cowan, 2000). Bioremediation is one of the most rapidly growing areas of environmental biotechnology. Use of bioremediation for environmental clean up is popular due to low costs and its public acceptability. Indeed, bioremediation stands to benefit greatly and advance even more rapidly with the adoption of molecular techniques developed originally for other areas of biotechnology. The 1990s was the decade of molecular microbial ecology (time of using molecular techniques in environmental biotechnology). Adoption of these molecular techniques made scientists realize that microbial populations in the natural environments are much more diverse than previously thought using traditional culture methods. Using molecular ecological methods, such as direct DNA isolation from environmental samples, denaturing gradient gel electrophoresis (DGGE), PCR methods, nucleic acid hybridization etc., we can now study microbial consortia relevant to pollutant degradation in the environment. These techniques promise to provide a better understanding and better control of environmental biotechnology processes, thus enabling more cost effective and efficient bioremediation of our toxic waste and contaminated environments.     

Current situation and challenges of specialized microbial resource centres in Russia

Establishment of national biological resource centres (BRCs) is of special concern and requires harmonization of regulations on microorganisms’ handling, improvement of legal control pertaining to intellectual property right, access to genetic resources and fair benefit sharing arising from their biotechnology application. As exemplified by the Regional Specialized Collection of Alkanotrophic Microorganisms (acronym IEGM, World Federation for Culture Collections # 768, ) hosted at the Institute of Ecology and Genetics of Microorganisms, the role of specialized microbial collections is emphasized as the governing factors of innovative development of biotechnology and bioindustry. The publication aims at drawing attention to the regional BRC being formed in the Perm Krai which provides the appropriate information on the holdings and is responsible for screening, study and maintenance of valuable microbial gene pool to meet the needs of ecology, industry and biotechnology, and for developing novel methodological approaches to studying extremotolerant microorganisms. This centre also contributes to the development and application of advanced achievements in enzymatic transformation of carbon compounds, production of fodder using non-traditional raw material, oil- and gas-prospecting activities, monitoring and bioremediation of contaminated sites.     

工业微生物在渗透胁迫下的应激反应及保护措施

发酵工业作为生物技术产业的重要组成部分,在我国工业结构中占据了极大比重,而在工业发酵后期菌体代谢物、中和剂以及补料物的累积使微生物受到极大的渗透胁迫,严重影响了细胞生长及目标产物代谢,致使发酵产量与效率偏低。本文主要针对高渗胁迫下微生物的细胞结构、应答途径、基因、蛋白、代谢、分裂机制进行综述与分析,并以微生物菌种特性结合工业发酵技术为改良思路,从菌种改良、外源添加保护剂、改良中和剂、去除渗透抑制因子、膜过滤技术等方面找寻潜在渗透保护措施,以期为发酵行业生产力水平的提升、节能减排降耗提供参考。     

Perspective of the Sugarcane Industry in Brazil

The sugarcane industry in Brazil is experiencing a rapid shift towards creating the grounds for a green and sustainable biorefinary industry. After 30 years of ProAlcool, the federal government program that boosted Brazil’s sugarcane industry by creating a mandate to blend ethanol with gasoline, flex fuel engines now dominate Brazil’s automobile industry. Currently, bioethanol replaces around 30% of the gasoline consumed in the country and its demand is projected to more that double in the next 10 years. On another front, the sugarcane genomics program created by FAPESP in the late 1990s paved the way for the establishment of innovative biotechnology startup companies that attracted the attention of the largest agro-biotechnology sector companies of the world. Almost all of these companies now have their sugarcane research centers surrounding the city of Campinas, São Paulo. In addition, innovative synthetic biology companies are developing technologies to produce diesel, jet fuel and other high value molecules using sugarcane juice as a carbon source. The sugarcane industry also teamed with petrochemical companies and already established operating plants to produce bioplastics. Innovations have also occurred in the field of co-generation of electricity from sugarcane bagasse. Currently sugarcane supplies 4% of the electricity needs of the country. Collectively, these innovations suggest that Brazil’s sugarcane industry could supply over 30% of the country energy needs by 2021 and a significant fraction of new bioproducts produced by its nascent biorefinary plants.     

Guidelines for establishing a culture collection within a biotechnology company

Culture collections within large pharmaceutical and biological companies act as depositories for preserving and maintaining important strains owned by, on the premises of, or sought on behalf of, the company. Other functions of a culture collection include: (a) regulating the flow of cultures into and out of the company; (b) recording and documenting such transfers; and (c) characterizing and/or identifying strains used for research, publications and patents. A culture collection within a biotechnology company is often required to maintain a diverse group of both prokaryote and eukaryote recombinant and non-recombinant strains. All strains and plasmids must be carefully characterized and preserved. A microbiologist with a strong background in microbial physiology, genetics and taxonomy is usually responsible for supervising the culture collection. This article focuses on guidelines for establishing a culture collection in biotechnology companies to serve the needs of both the scientist and the company.     

Preparation and properties of HDPE/sugarcane bagasse cellulose composites obtained for thermokinetic mixer

The use of natural fibers as reinforcement for thermoplastics has generated much interest due to their low cost, possibility of environmental protection and use of locally available renewable resources. In this work the mechanical and morphological properties of high density polyethylene/pre-treated and modified residues from sugarcane bagasse cellulose composites were analyzed. Composites were produced by a thermokinetic mixer. The microstructural analyses of fracture surface from composites can be easily evaluated by microscopic techniques. Results showed that the modification of sugarcane bagasse cellulose with zirconium oxychloride was successfully accomplished and that this reinforcement material with high density polyethylene showed tensile strength higher than non-modified sugarcane bagasse cellulose. Modification in the sugarcane bagasse cellulose influenced directly in mechanical properties of the composite material. This can be observed by the fracture surface, which showed that modified cellulose sugarcane bagasse improved interfacial adhesion between fiber and matrix.     

Increased digestibility of bagasses by pretreatment with alkalis and steam explosion

Alkali treatment and steam explosion of bagasse were investigated in order to develop economical and effective methods of increasing the digestibility of bagasse. The treated bagasse was to be used as a substrate for the production of volatile fatty acids by anaerobic acidogenic bacteria. The alkalis examined were NaOH, NH(3) (aqueous), NaOH + NH(3), Ca(OH)(2), and Ca(OH)(2) + Na(2)CO(3), at ambient temperature and in combination with steam explosion at 200 degrees C, 6.9 MPa, and 5 min cooking times. Digestibilities of up to 733 g organic matter (OM)/kg bagasse dry matter (DM) were obtained for bagasse treated with NaOH and Ca(OH)(2) + Na(2)CO(3); less than 430 g OM was obtained for bagasse treated with aqueous NH(3); and up to 724 g OM was obtained for bagasse treated with Ca(OH)(2). This digestibility was only achieved by using high concentrations of Ca(OH)(2), i.e., 180-300 g/kg bagasse. Steam explosion increased the digestibility of bagasse up to 740 g OM in the presence of alkali but only to 610 g OM in the absence of alkali. The digestibility of bagasse without pretreatment was 190 g OM/kg bagasse DM. More than one-half the hemicellulose present was solubilized by pretreatment. The composition of the liquid fraction of steam-exploded material was examined and contained mainly xylose monomers and oligomers (112 g/kg original bagasse DM) and acetic acid (33 g/kg original DM). The relative costs of the alkalis used were obtained for the United States, Australia, and Europe. Lime [Ca(OH)(2)] was the least expensive alkali per unit of additional digestible OM obtained. Ammonia was the most expensive alkali to use, except in the United States where the difference in its cost relative to other alkalis was smaller. However, ammonia provides organic nitrogen for microbial growth, and could be recycled. With acidogenic fermentations, alkali is able to double as a neutralizing agent during fermentation. Thus, concentrations of alkali equal to that required for neutralization may be used in pretreatment. Concentrations of Ca(OH)(2) as high as 300 g/kg bagasse were needed for neutralization and should, therefore, be considered for pretreatment. Steam explosion of bagasse resulted in digestible, sterilized substrates of small particle size with readily separable liquid and pulp streams.     

Use of sugarcane bagasse as an alternative low-cost support material during the rooting stage of apple micropropagation

Summary Studies were carried out to evaluate sugarcane bagasse as an alternative to agar for micropropagation of apple clones to reduce the cost of micropropagation and improve the quality of the propagules. Significant improvement in the in vitro rooting process, coupled with cost reduction, were obtained by the use of sugarcane bagasse as a substitute for the traditionally used agar-gelled medium. The tests were undertaken with micro-cuttings of the apple rootstock Marubakaido (Malus prunifolia Borkh.) using a rooting medium composed of half-strength Murashige and Skoog salts and vitamins, 3% (w/v) sucrose, and 0.49 μM indole-3-butyric acid. The plants grown on sugarcane bagasse yielded a 22% increase in root length, 20% increase in plant length, and 63% increase in the number of roots, compared with agar-grown micro-cuttings. Particle size of the sugarcane bagasse had a significant impact on all those parameters, and the best results were obtained with bagasse comprising particles smaller than 0.18 mm. The results demonstrated that the sugarcane bagasse could be used effectively as a substitute for agar during rooting of apple shoots.     

Microbial synthesis of chalcogenide semiconductor nanoparticles: a review

Chalcogenide semiconductor quantum dots are emerging as promising nanomaterials due to their size tunable optoelectronic properties. The commercial synthesis and their subsequent integration for practical uses have, however, been contorted largely due to the toxicity and cost issues associated with the present chemical synthesis protocols. Accordingly, there is an immediate need to develop alternative environment‐friendly synthesis procedures. Microbial factories hold immense potential to achieve this objective. Over the past few years, bacteria, fungi and yeasts have been experimented with as eco‐friendly and cost‐effective tools for the biosynthesis of semiconductor quantum dots. This review provides a detailed overview about the production of chalcogen‐based semiconductor quantum particles using the inherent microbial machinery.     

Use of sugarcane bagasse and grass hydrolysates as carbon sources for xylanase production by Bacillus circulans D1 in submerged fermentation

The use of sugarcane bagasse and grass as low cost raw material for xylanase production by Bacillus circulans D1 in submerged fermentation was investigated. The microorganism was cultivated in a mineral medium containing hydrolysate of bagasse or grass as carbon source. High production of enzyme was obtained during growth in media with bagasse hydrolysates (8.4 U/mL) and in media with grass hydrolysates (7.5 U/mL). Xylanase production in media with hydrolysates was very close to that obtained in xylan containing media (7.0 U/mL) and this fact confirm the feasibility of using this agro-industrial byproducts by B. circulans D1 as an alternative to save costs on the enzyme production process.     

Bioethanol and biodiesel: Alternative liquid fuels for future generations

The global population is expected to increase by approximately 3 billion people by 2050. With this increase in population, industry, transportation the cost of fossil fuels will grow dramatically. New technologies are needed for fuel extraction using feedstocks that do not threaten food security, cause minimal or no loss of natural habitat and soil carbon. At the same time, waste management has to be improved and environmental pollution should be minimized or eliminated. Liquid biofuels such as lignocellulosic‐based ethanol from plant biomass and algal‐based biodiesel are sustainable, alternative biofuels that could stabilize national security and provide clean energy for future generations. Ideally, the technology should also foster recycling of agricultural feedstocks and improve soil fertility and human health. This article provides updated information on the energy potential and breadth of liquid biofuel biotechnology.     

Proteins identified through predictive metagenomics as potential biomarkers for the detection of microbiologically influenced corrosion

The unpredictability of microbial growth and subsequent localized corrosion of steel can cause significant cost for the oil and gas industry, due to production downtime, repair, and replacement. Despite a long tradition of academic research and industrial experience, microbial corrosion is not yet fully understood and thus not effectively controlled. In particular, biomarkers suitable for diagnosing microbial corrosion which abstain from the detection of the classic signatures of sulfate-reducing bacteria are urgently required. In this study, a natural microbial community was enriched anaerobically with carbon steel coupons and in the presence of a variety of physical and chemical conditions. With the characterization of the microbiome and of its functional properties inferred through predictive metagenomics, a series of proteins were identified as biomarkers in the water phase that could be correlated directly to corrosion. This study provides an opportunity for the further development of a protein-based biomarker approach for effective and reliable microbial corrosion detection and monitoring in the field.