Recent Patents on Biotechnology (v.10, #1)

Meet Our Editorial Board Member: by William C.S. Cho (1-1).

Preface by Nikolaos E. Labrou (2-2).

Plant Volatile Genomics: Recent Developments and Putative Applications in Agriculture by Ishita Paul, Pratapbhanu Singh Bhadoria, Adinpunya Mitra (4-11).
The review of patents reveals that investigation of plant volatiles and their biosynthetic pathways is a relatively new field in plant biochemistry. The diversity of structure and function of these volatiles is gradually being understood. However, the great diversity of volatile biochemicals plants emit through different parts plays numerous roles in stress resistance and other ecological interactions. From an agronomic point of view, regulation volatile production in crop plants may lead to desirable changes in plant defence, pollinator attraction and post-harvest qualities. In several crop species, genetic manipulation or metabolic channelling have led to altered emission I aroma profiles. This short review summarizes some recent cases of artificial manipulation of volatile profile in planta or in transformed microbial systems.

Molecular Genetic Approaches for Environmental Stress Tolerant Crop Plants: Progress and Prospects by Ranjeet Kaur, Rupam Kumar Bhunia, Ananta Kumar Ghosh (12-29).
Background: Global food security is threatened by the severe environmental conditions that have reduced the worldwide crop yield. Plants possess inherent mechanisms to cope with the initial stress phase but to ensure their survival through harsh climate, the intervention of genetic engineering is desirable.
Objective: We present a comprehensive review on the progress made in the field of developing environmental stress tolerant crops and the prospects that can be undertaken for achieving it.
Methods: We review the effects of abiotic and biotic stresses on crop plants, and the use of different molecular genetic approaches to cope with these environmental stresses for establishment of sustainable agriculture. The various strategies employed in different crops have also been discussed. We also summarized the major patents in the field of plant stress tolerance that have been granted in the last five years.
Results: On the basis of these analyses, we propose that genetic engineering of crops is the preferred approach over the traditional methods for yielding healthier and viable agriculture in response to the different stressful environments. The wild progenitors of cultivated crop species can prove to be highly potential genetic resources in this regard and can be exploited to produce better crops that are relatively tolerant towards various environmental stresses.
Conclusion: Thus, elucidation of genetic loci and deciphering the underlying mechanisms that confer tolerance to plants against stressful conditions followed by its successful introgression into elite, high-yielding crop varieties can be an effective way to engineer the crops for sustainable agriculture.

Recent Inventions and Trends in Algal Biofuels Research by Ankush Karemore, Manoranjan Nayak, Ramkrishna Sen (30-42).
Background: In recent times, when energy crisis compounded by global warming and climate change is receiving worldwide attention, the emergence of algae, as a better feedstock for third-generation biofuels than energy crops or plants, holds great promise. As compared to conventional biofuels feedstocks, algae offer several advantages and can alone produce a significant amount of biofuels sustainably in a shorter period to fulfill the rising demand for energy.
Objective: Towards commercialisation, there have been numerous efforts put for- ward for the development of algae-derived biofuel. This article reviews and summarizes the recent inventions and the current trends that are reported and captured in relevant patents pertaining to the novel methods of algae biomass cultivation and processing for biofuels and value-added products. In addition, the recent advancement in techniques and technologies for microalgal biofuel production has been highlighted.
Methods: Various steps involved in the production of algal biofuels have been considered in this article. Moreover, the work that advances to improve the efficiency and cost-effectiveness of the processes for the manufacture of biofuels has been presented. Our survey was conducted in the patent databases: WIPO, Spacenet and USPTO.
Results: There are still some technological bottlenecks that could be overcome by designing advanced photobioreactor and raceway ponds, developing new and low cost technologies for biomass cultivation, harvesting, drying and extraction.
Conclusion: Recent advancement in algae biofuels methods is directed toward developing efficient and integrated systems to produce biofuels by overcoming the current challenges. However, further research effort is required to scale-up and improve the efficiency of these methods in the upstream and downstream technologies to make the cost of biofuels competitive with petroleum fuels.

Food Biotechnology: A Step Towards Improving Nutritional Quality of Food for Asian Countries by Archana Dash, Debajyoti Kundu, Mohan Das, Debalina Bose, Sunita Adak, Rintu Banerjee (43-57).
Background: Food-based technologies established itself as a vast, inter-disciplinary and multifaceted research area are at crossroads of scientific and technological advancements. The review of patents reveals that growing concerns over the source of nourishment for the burgeoning population, its quality, quantity and safety, along with associated human and environmental welfare has inspired global researchers to implement several new bio-based technologies.
Objective: Biotechnological interventions in food sector have been aimed at enhancing/modifying taste, aroma, shelf-life, texture and nutritional value of food products employing fermentation, enzyme technology, nanotechnology and molecular biology. The use of whole microbes as a source of nutrition and genetically modified microorganisms to be used as food or genetically modified food have been successfully attempted, which has addressed the mass population and malnutrition. Further, the processing techniques have been improved along with the proper utilization of food wastes for the generation of many useful byproducts.
Conclusion: The article covers broadly the impact of biotechnological interventions in food sector. Techniques mostly discussed include not only the nutrient enriched food production but also byproduct utilization through proper improvisation of food waste into bioenergy, biomanure and other value added products, which is of great economic and environmental importance. This article reviews the overall aspects in relation to some of the recent advancements in food sector.

Enzymes- An Existing and Promising Tool of Food Processing Industry by Lalitagauri Ray, Sunita Pramanik, Debabrata Bera (58-71).
Background: The enzyme catalyzed process technology has enormous potential in the food sectors as indicated by the recent patents studies. It is very well realized that the adaptation of the enzyme catalyzed process depends on the availability of enzyme in affordable prices.
Objective: Enzymes may be used in different food sectors like dairy, fruits & vegetable processing, meat tenderization, fish processing, brewery and wine making, starch processing and many other. Commercially only a small number of enzymes are used because of several factors including instability of enzymes during processing and high cost.
Method: More and more enzymes for food technology are now derived from specially selected or genetically modified microorganisms grown in industrial scale fermenters. Enzymes with microbial source have commercial advantages of using microbial fermentation rather than animal and plant extraction to produce food enzymes.
Conclusion: At present only a relatively small number of enzymes are used commercially in food processing. But the number is increasing day by day and field of application will be expanded more and more in near future. The purpose of this review is to describe the practical applications of enzymes in the field of food processing.

Immunomodulatory and Anti-cancer Properties of Pharmacologically Relevant Mushroom Glycans by Kangabam Sanjana Priyadarshini Devi, Tapas Kumar Maiti (72-78).
The review of patents reveals that for decades, mushrooms have been regarded as an ethnic medicine and widely used in the treatment of various life-threatening diseases. One of the most important bioactive compounds isolated from these mushrooms are polysaccharides or glycans which exhibit potential immunomodulatory and anticancer properties. The immune stimulating properties of these glycans are linked to their origin, structure, composition, water solubility and conformations in solution. Moreover, modification of glycans to increase its size to a particulate form has also shown significant increase in its activity. Glycans are regarded as pathogen associated molecu- lar pattern (PAMP) like molecules which bind to specific pattern recognition receptors (PRRs) on the surface of immune cells. This binding induces the activation of a signaling cascade that directs the synthesis of a repertoire of immune effector molecules. Anticancer effects of glycans have been mostly associated with an increased immune activity while few cases are available which advocate its direct role on cancer cells. This review thereby provides a comprehensive outlook on the structure-function relationship of these pharmacologically relevant mushroom glycans.

Bio-electricity Generation using Jatropha Oil Seed Cake by Hifjur Raheman, Debasish Padhee (79-85).
Background: The review of patents reveals that Handling of Jatropha seed cake after extraction of oil is essential as it contains toxic materials which create environmental pollution.
Methods: The goal of this work is complete utilisation of Jatropha seeds. For this purpose, Jatropha oil was used for producing biodiesel and the byproduct Jatropha seed cake was gasified to obtain producer gas. Both biodiesel and producer gas were used to generate electricity. To achieve this, a system comprising gasifier, briquetting machine, diesel engine and generator was developed.
Results: Biodiesel was produced successfully using the method patented for biodiesel production and briquettes of Jatropha seed cake were made using a vertical extruding machine. Producer gas was obtained by gasifying these briquettes in a downdraft gasifier. A diesel engine was then run in dual fuel mode with biodiesel and producer gas instead of only diesel. Electricity was generated by coupling it to a generator.
Conclusion: The cost of producing kilowatthour of electricity with biodiesel and diesel in dual fuel mode with producer gas was found to be 0.84 $ and 0.75 $, respectively as compared to 0.69 $ and 0.5 $ for the same fuels in single fuel mode resulting in up to 48 % saving of pilot fuel. Compared to singlefuel mode, there was 25-32 % reduction in system and brake thermal efficiency along with significantly lower NOx, higher CO and CO2 emissions when the bio-electricity generating system was operated in dual fuel mode. Overall, the developed system could produce electricity successfully by completely uti- lising Jatropha seeds without leaving any seed cake to cause environmental pollution.

Background: Antimicrobial potency of herbal extracts is well known. The review of patents and research articles revealed that several herbal extracts have been employed in the formulation of topical products such as creams, exclusive of the cream reported in the present study. 0ur previous study has established antimicrobial potency of supercritical carbon dioxide extracts of tuberose flowers, better known for its sweet fragrance.
Objective: The present work focuses on formulating a topical antimicrobial herbal cream with methyl eugenol (principal antimicrobial compound) rich - supercritical carbon dioxide extract of tuberose flowers, having good combination of phytochemical and antimicrobial potencies.
Methods: Supercritical carbon dioxide parameters such as temperature, pressure and time were optimized using full factorial experimental design to obtain methyl eugenol-rich extracts. A cream was formulated using the extract having the best combination of phytochemical and antimicrobial potencies and was assayed further for in vitro antimicrobial potency; physiochemical and sensory properties. Two commercial antimicrobial cream samples were used as reference samples in the study.
Result: The extract obtained at 40°C, 10 MPa, 135 min at 1 L min-1 flow rate of gaseous C02 showed the best combination of phytochemical and antimicrobial potencies and was used for formulation of herbal creams. The cream formulated with 5% w/w of extract arrested growth of the common human skin pathogen Staphylococcus aureus and showed stable physiochemical properties and high sensory appeal for a year.
Conclusion: The cream could be considered as a 'finished herbal product' in compliance with the World Health 0rganization guidelines.

Effect of Packaging on Shelf-life and Lutein Content of Marigold (Tagetes erecta L.) Flowers by Sayani Pal, Probir Kumar Ghosh, Paramita Bhattacharjee (103-120).
Background: African marigold (Tagetes erecta L.) flowers are highly valued for their ornamental appeal as well as medicinal properties. However, their short shelf lives cause high post-harvest loss and limit their export potential. The review of patents and research articles revealed that different types of packaging designs/materials have been successfully employed for extension of shelf lives of cut flowers.
Objective: The current work focuses on designing of different packaging configurations and selection of best configuration for preservation of marigold cut flowers.
Methods: Ten packaging configurations, composed of four different packaging materials i.e., low density polyethylene (LDPE), polyethylene terephthalate, glassine paper and cellophane paper, were designed. Each pack, consisting of 20 ± 1 g of marigold flowers along with non-packaged control set were stored at 23 ± 2°C, 80% R.H., in an environmental chamber and the flowers were evaluated for their sensory attributes, phytochemical characteristics and physicochemical parameters of senescence to determine their shelf lives.
Results: Flowers packed in LDPE bag showed highest shelf life of 8 days with a lead of 4 days compared to control (shelf life - 4 days). This study also established for the first time the phenomenon of carotenogenesis in marigold cut flowers with significantly (P<0.01) higher production of lutein in LDPE packaged flowers.
Conclusion: LDPE pack was the best design among the ten package designs, in preserving lutein content of marigold flowers and extending their shelf lives. This economically viable packaging can not only boost the export potential of this ornamental flower, but also allow utilization of nutraceutical potency of lutein.

Background: Penicillin G amidase (PGA) (EC are enzymes that are mainly involved in the synthesis of semi-synthetic }-lactam antibiotics. Soluble PGA is costly and lacks long term operational stability. We revised most of the patents related to Penicillin G amidase (PGA) immobilization in the section "Recent Patents on Immobilized Penicillin G Amidase".
Objective: The aim of this work was to study comparative biochemical property of PGA enzyme immobilized in two hydro-gel beads - Ca-alginate and alginate+chitosan hybrid and morphologically characterised by SEM.
Methods: PGA immobilized in alginate+chitosan hybrid bead shows high pH and thermal stability. Km, Vmax and Effectiveness factor (1) value of free PGA were 56.19 mg/ml, 1.786 U/ml and 1, respectively. These parameters for PGA immobilized alginate beads were 64.84 mg/ml, 0.781U/ml and 0.437, respectively and for PGA immobilized alginate+chitosan hybrid beads were 87.08 mg/ml, 0.622 U/ml and 0.348, respectively.
Results: Immobilized PGA on alginate+chitosan hybrid beads gave the highest thermal stability, reusability and storage stability than alginate immobilized PGA. Conclusion: The entrapment of PGA on alginate+chitosan hybrid beads revealed several advantages and could be used in 6APA (6- aminopenicillanic acid) production.

Background: Pseudomonas aeruginosa is one of the potent opportunistic pathogens associated with respiratory and urinary tract infection. The bacterium owes its pathogenicity due to the intrinsic resistance to antibiotics and disinfectants.
Objective: The present study is focused on the synthesis of antibacterial chitosan coated iron oxide nanoparticles for rapid inactivation of Pseudomonas aeruginosa. We have discussed the relevant patents on synthesis and antibacterial potential of metallic nanoparticles and chitosan.
Method: Chitosan coated iron oxide nanoparticles were synthesized by coprecipitation method at room temperature using non-toxic chitosan and iron salts in alkali media. The particles were characterized and evaluated for antibacterial property against Pseudomonas aeruginosa.
Results: The average size of the particles was measured as 52 nm. The surface area of the coated particles was as high as 90 ±5 m2/g. FTIR spectra confirmed the coating of chitosan on nanoparticles. The coated particles showed excellent antibacterial activity against the bacteria. The minimum inhibitory concentration of the coated particles was 105).g mol-1. The morphological alteration and cytoplasmic leakage of bacteria were confirmed by SEM image and release of intracellular constituents, respective- ly. Higher 260 nm absorbance value confirmed stronger antibacterial activity of the coated nanoparticles as compared to pure chitosan and bare iron oxide nanoparticles.
Conclusion: The study indicated that chitosan coated iron oxide nanoparticles have superior antibacterial property as compared to pure chitosan and iron oxide nanoparticles.

Patent Selections: (140-141).

ERRATUM (142-142).